{"gene":"MAK","run_date":"2026-04-28T18:30:27","timeline":{"discoveries":[{"year":1990,"finding":"MAK (male germ cell-associated kinase) encodes a novel serine/threonine protein kinase whose kinase domain shares ~40% identity with the cdc2-CDC28 gene family; its transcripts are expressed almost exclusively in testicular germ cells entering and after meiosis.","method":"cDNA cloning, sequence analysis, Northern blot","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1 — original cloning with sequence analysis establishing kinase identity and expression pattern, foundational paper with 113 citations","pmids":["2183027"],"is_preprint":false},{"year":1993,"finding":"MAK protein products (66 and 60 kDa) are most abundant in late pachytene spermatocytes, localize as soluble cytoplasmic proteins, associate with a 210 kDa cellular protein, and phosphorylate this 210 kDa protein on serine and threonine residues both in vitro (immunoprecipitate kinase assay) and in vivo (32Pi labeling).","method":"Subcellular fractionation, immunoprecipitation kinase assay, [35S]methionine labeling, 32Pi metabolic labeling","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1-2 — in vitro kinase assay combined with in vivo phosphorylation and fractionation, replicated with multiple antibodies","pmids":["8321219"],"is_preprint":false},{"year":2006,"finding":"MAK and MRK/ICK require dual phosphorylation in a TDY activation-loop motif: threonine phosphorylation (T157 in MRK) is catalyzed by CCRK (cell cycle-related kinase), while tyrosine phosphorylation is by autophosphorylation. Protein phosphatase 5 (PP5) dephosphorylates MRK at T157, acting as the opposing regulator. MRK preferentially phosphorylates R-P-X-S/T-P motifs (consensus determined by combinatorial peptide library screening), and phosphorylates the antiapoptotic protein Scythe at T1080.","method":"In vitro kinase assay, combinatorial peptide library screening, site-directed mutagenesis, mass spectrometry, co-immunoprecipitation","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1 — reconstituted kinase cascade in vitro with mutagenesis and MS validation, multiple orthogonal methods in single study","pmids":["16954377"],"is_preprint":false},{"year":2010,"finding":"Mak localizes to the connecting cilia and outer-segment axonemes of photoreceptor cells and negatively regulates ciliary length. In Mak-null retinas, photoreceptors show elongated cilia with accumulated IFT88, IFT57, expanded Kif3a, and acetylated α-tubulin signals, and rhodopsin accumulates abnormally in cell bodies. Overexpression of RP1 (a microtubule-associated protein) induces ciliary elongation, and Mak coexpression rescues this; the RP1 N-terminal portion is phosphorylated by Mak in vitro, linking Mak kinase activity to ciliary length control.","method":"Mak-null mouse model, immunohistochemistry, in vitro kinase assay, overexpression rescue experiments","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — genetic KO with defined cellular phenotype, in vitro kinase assay identifying RP1 as substrate, rescue experiments; 117 citations","pmids":["21148103"],"is_preprint":false},{"year":2011,"finding":"Missense mutations in MAK that alter conserved kinase residues abolish kinase activity as demonstrated by in vitro kinase assay, establishing that MAK kinase activity is essential for its function in photoreceptor ciliary maintenance.","method":"In vitro kinase assay with disease-causing mutants","journal":"American journal of human genetics","confidence":"High","confidence_rationale":"Tier 1 — in vitro kinase assay directly linking catalytic activity to human disease mutations","pmids":["21835304"],"is_preprint":false},{"year":2011,"finding":"MAK is expressed in the inner segments, cell bodies, and axons of rod and cone photoreceptors in human donor tissue. An exon 9-containing MAK isoform (also including a novel exon 12) is the predominant transcript in retinal precursor cells, and failure to switch to this isoform (caused by an Alu insertion in exon 9) results in loss of normal MAK protein and retinitis pigmentosa.","method":"Immunohistochemistry of human donor tissue, iPSC differentiation into retinal precursors, RT-PCR isoform analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — direct localization in human tissue plus functional isoform analysis in iPSC-derived retinal precursors; 177 citations","pmids":["21825139"],"is_preprint":false},{"year":2002,"finding":"MAK-deficient (Mak-/-) mice are viable and fertile with intact spermatogenesis, demonstrating that MAK is not essential for sperm formation, though Mak-/- males show mildly reduced litter sizes and reduced sperm motility in vitro.","method":"Targeted gene knockout, fertility and spermatogenesis phenotyping","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — clean genetic KO with defined phenotypic readouts in multiple assays","pmids":["11971961"],"is_preprint":false},{"year":2019,"finding":"In C. elegans, DYF-18/CCRK and DYF-5/MAK act in a kinase cascade to regulate ciliary length and branching morphology: loss of dyf-18 or dyf-5 results in dramatically elongated, unbranched AWA olfactory neuron cilia. The mechanism involves regulation of axonemal microtubule stability, as EBP-2 end-binding protein decorates axonemal microtubules along their lengths, tubulin load increases, and tubulin turnover is reduced in dyf-18 mutants. Microtubule-destabilizing tubulin mutations and IFT protein mutations that disrupt tubulin transport suppress AWA cilia elongation in dyf-18 mutants.","method":"C. elegans genetic epistasis, live imaging, IFT motor localization, genetic suppressor analysis","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 1-2 — genetic epistasis with multiple suppressor mutations plus live imaging defining mechanism","pmids":["30955935"],"is_preprint":false},{"year":2022,"finding":"The ciliary kinase DYF-5/MAK phosphorylates multiple sites within the tubulin-binding module of IFT-74, reducing its tubulin-binding affinity approximately sixfold and promoting tubulin unloading at the ciliary tip. Ablation or constitutive activation of IFT-74 phosphorylation abnormally elongates or shortens sensory cilia in C. elegans neurons, respectively.","method":"In vitro phosphorylation assay, affinity binding measurements, C. elegans genetic analysis with phosphomimetic/phosphoablating IFT-74 mutations","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution of phosphorylation event with quantified affinity change, validated in vivo with gain/loss-of-function mutations","pmids":["35969738"],"is_preprint":false},{"year":2024,"finding":"MAK localizes to the ciliary tip and functions as an IFT regulator cooperatively with ICK. Simultaneous disruption of Mak and Ick in mice results in loss of photoreceptor ciliary axonemes and severe retinal degeneration. CCRK kinase is identified as an upstream activator of both Mak and Ick in retinal photoreceptor cells. Gene delivery of Ick and pharmacological inhibition of FGFR (a negative regulator of Ick) ameliorate retinal degeneration in Mak-/- mice.","method":"Double-knockout mouse model, live imaging, subretinal gene delivery, pharmacological FGFR inhibition","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 2 — genetic double-KO with defined phenotype, pathway placement via epistasis, therapeutic rescue validates mechanism","pmids":["39293864"],"is_preprint":false},{"year":2021,"finding":"Viral overexpression of the retinal MAK transgene in patient iPSC-derived photoreceptor precursor cells restores MAK transcript and protein. Overexpression of retinal MAK in patient-derived fibroblasts and mak-knockdown zebrafish restores the ability to regulate primary cilia length, and mitigates the visual defect in mak knockdown zebrafish.","method":"iPSC-derived cells, viral gene delivery, zebrafish morpholino knockdown, cilia length assay","journal":"Gene therapy","confidence":"Medium","confidence_rationale":"Tier 2 — functional rescue of cilia length defect in patient cells and zebrafish model, but single lab study","pmids":["34518651"],"is_preprint":false},{"year":2022,"finding":"PCARE contains evolutionary conserved binding sites for the photoreceptor kinase MAK at its C-terminal region. Deletion of the MAK-binding motifs in PCARE did not abolish ciliary localization but affected downstream WASF3-mediated ciliary tip membrane expansion, suggesting MAK interacts with PCARE to regulate actin-driven photoreceptor outer segment disk formation.","method":"Co-expression studies, deletion mutagenesis, ciliary expansion assay","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 3 — functional co-expression with deletion mutagenesis, but direct binding not biochemically confirmed with pulldown","pmids":["35253837"],"is_preprint":false},{"year":2025,"finding":"In Chlamydomonas, CrMAK (ortholog of mammalian ICK/MAK) localizes to cilia and is enriched at the ciliary tip during active ciliary assembly. CrMAK associates with the axoneme via its C-terminal tail, acts downstream of the LF1/LF2/LF3 ciliary length-regulatory complex, and phosphorylates FAP256/CEP104 (a microtubule plus-end tracking protein). Loss of CrMAK or its kinase activity leads to aciliated cells, and dephosphorylation of FAP256/CEP104 impairs ciliary assembly.","method":"Chlamydomonas mutant analysis, in vitro kinase assay, co-immunoprecipitation, live imaging, mutagenesis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution of phosphorylation with identified substrate (FAP256/CEP104), genetic epistasis placing MAK downstream of LF complex, loss-of-function phenotype","pmids":["41231942"],"is_preprint":false}],"current_model":"MAK (male germ cell-associated kinase) is a serine/threonine kinase activated by CCRK-mediated phosphorylation of its TDY activation-loop threonine, that localizes to the ciliary tip where it phosphorylates IFT-74 (reducing tubulin-binding affinity to promote tubulin unloading), RP1/microtubule-associated proteins (to limit axonemal extension), and FAP256/CEP104 (to regulate axonemal microtubule assembly), thereby acting as a master negative regulator of ciliary length whose loss causes progressive photoreceptor degeneration and retinitis pigmentosa."},"narrative":{"teleology":[{"year":1990,"claim":"Identification of MAK as a novel CDC2-related serine/threonine kinase expressed predominantly in meiotic germ cells established the gene's kinase identity and pointed to a specialized role in differentiated cell types.","evidence":"cDNA cloning, sequence analysis, and Northern blot in mouse testis","pmids":["2183027"],"confidence":"High","gaps":["No substrate or pathway identified","Function in non-testicular tissues unknown"]},{"year":1993,"claim":"Demonstration that MAK associates with and phosphorylates a 210 kDa protein in spermatocytes both in vitro and in vivo established that MAK functions as an active kinase with at least one endogenous substrate in germ cells.","evidence":"Immunoprecipitate kinase assay and 32Pi metabolic labeling in pachytene spermatocytes","pmids":["8321219"],"confidence":"High","gaps":["Identity of the 210 kDa substrate unknown","Physiological role of this phosphorylation unclear"]},{"year":2002,"claim":"The finding that Mak-knockout mice are viable and fertile with only mild sperm motility defects redirected the field away from an essential spermatogenesis role and opened the question of MAK's primary physiological function.","evidence":"Targeted gene knockout in mice with fertility and spermatogenesis phenotyping","pmids":["11971961"],"confidence":"High","gaps":["Non-testicular phenotypes not yet examined","Functional redundancy with ICK not tested"]},{"year":2006,"claim":"Reconstitution of the CCRK→MAK/MRK activation cascade, including identification of the TDY activation-loop mechanism, PP5 as opposing phosphatase, and the R-P-X-S/T-P substrate consensus, defined how MAK is activated and what it recognizes.","evidence":"In vitro kinase assay, combinatorial peptide library screening, mutagenesis, mass spectrometry","pmids":["16954377"],"confidence":"High","gaps":["Physiological substrates matching R-P-X-S/T-P consensus not identified in vivo","Whether CCRK activates MAK in all tissues unknown"]},{"year":2010,"claim":"Discovery that Mak localizes to photoreceptor connecting cilia and negatively regulates ciliary length — with Mak-null retinas showing elongated cilia, IFT accumulation, and rhodopsin mislocalization — fundamentally recast MAK as a ciliary kinase, with RP1 identified as a direct phospho-substrate linking kinase activity to axonemal length control.","evidence":"Mak-null mouse model, immunohistochemistry, in vitro kinase assay, overexpression rescue","pmids":["21148103"],"confidence":"High","gaps":["Whether RP1 phosphorylation alone accounts for length phenotype unknown","Mechanism of IFT accumulation not defined"]},{"year":2011,"claim":"Two studies linked MAK loss-of-function to human retinitis pigmentosa: kinase-dead missense mutations were shown to abolish catalytic activity, and an Alu insertion disrupting retinal MAK isoform splicing was identified as a disease-causing allele, establishing MAK as a retinitis pigmentosa gene requiring kinase activity for photoreceptor survival.","evidence":"In vitro kinase assay with patient mutations; iPSC-derived retinal precursor isoform analysis; immunohistochemistry of human donor retina","pmids":["21835304","21825139"],"confidence":"High","gaps":["Natural history and genotype-phenotype correlations incompletely characterized","Whether cone versus rod photoreceptors are differentially affected unclear"]},{"year":2019,"claim":"Genetic epistasis in C. elegans demonstrated that the CCRK→MAK cascade controls ciliary length by regulating axonemal microtubule stability and tubulin delivery via IFT, with microtubule-destabilizing mutations and IFT mutations suppressing the elongation phenotype.","evidence":"C. elegans genetic epistasis, live imaging, suppressor analysis in AWA olfactory neurons","pmids":["30955935"],"confidence":"High","gaps":["Direct phosphorylation target mediating microtubule stability not identified in this study","Whether branching versus length regulation are separable unknown"]},{"year":2021,"claim":"Viral delivery of retinal MAK rescued cilia length defects in patient iPSC-derived photoreceptor precursors and visual function in mak-knockdown zebrafish, providing proof-of-concept for gene therapy.","evidence":"iPSC-derived cells, viral gene delivery, zebrafish morpholino knockdown, cilia length assay","pmids":["34518651"],"confidence":"Medium","gaps":["Long-term efficacy and safety of gene delivery not assessed","Single-lab study awaits independent replication"]},{"year":2022,"claim":"Identification of IFT-74 as a direct MAK/DYF-5 substrate whose phosphorylation reduces tubulin-binding affinity ~6-fold provided a molecular mechanism for how MAK promotes tubulin unloading at the ciliary tip to control axonemal length.","evidence":"In vitro phosphorylation assay, affinity measurements, phosphomimetic/phosphoablating IFT-74 mutations in C. elegans","pmids":["35969738"],"confidence":"High","gaps":["Whether IFT-74 phosphorylation operates identically in mammalian photoreceptors not shown","Contribution of IFT-74 phosphorylation relative to RP1 phosphorylation unknown"]},{"year":2022,"claim":"PCARE was identified as a MAK-interacting protein at the ciliary tip, with MAK-binding motifs required for WASF3-mediated membrane expansion, suggesting MAK regulates actin-driven photoreceptor outer segment disk formation.","evidence":"Co-expression studies, deletion mutagenesis, ciliary expansion assay","pmids":["35253837"],"confidence":"Medium","gaps":["Direct biochemical binding not confirmed by pulldown or co-IP","Whether MAK phosphorylates PCARE not tested","Single co-expression system without in vivo validation"]},{"year":2024,"claim":"Double knockout of Mak and Ick in mice revealed functional cooperativity between these kinases, with combined loss causing complete loss of photoreceptor ciliary axonemes and severe degeneration; Ick gene delivery and pharmacological FGFR inhibition rescued Mak-null retinal degeneration, establishing therapeutic redundancy.","evidence":"Double-knockout mouse model, subretinal gene delivery, pharmacological FGFR inhibition","pmids":["39293864"],"confidence":"High","gaps":["Whether MAK and ICK share identical substrates or have distinct targets not resolved","Optimal therapeutic window for intervention not defined"]},{"year":2025,"claim":"In Chlamydomonas, CrMAK was placed downstream of the LF1/LF2/LF3 length-regulatory complex and shown to phosphorylate FAP256/CEP104, a microtubule plus-end tracking protein, whose dephosphorylation impairs ciliary assembly — adding a third identified substrate to MAK's ciliary repertoire.","evidence":"Chlamydomonas mutant analysis, in vitro kinase assay, co-immunoprecipitation, live imaging","pmids":["41231942"],"confidence":"High","gaps":["Whether mammalian CEP104 is similarly regulated by MAK not tested","Structural basis for CrMAK axonemal association via C-terminal tail unresolved"]},{"year":null,"claim":"The relative contributions of MAK's multiple substrates (IFT-74, RP1, FAP256/CEP104, PCARE) to ciliary length control and photoreceptor outer segment homeostasis remain unintegrated, and no structural model of MAK exists to explain substrate selectivity or the cooperative relationship with ICK.","evidence":"","pmids":[],"confidence":"High","gaps":["No crystal or cryo-EM structure of MAK","Quantitative contribution of each substrate to ciliary length phenotype not parsed","Whether MAK has non-ciliary functions in retinal or other neurons remains open"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,2,3,4,8,12]}],"localization":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[3,5,9,12]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[3,7,8,9,12]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[3,7,8,9,12]},{"term_id":"R-HSA-9709957","term_label":"Sensory Perception","supporting_discovery_ids":[3,5,9,10]}],"complexes":[],"partners":["CCRK","ICK","RP1","IFT74","PCARE","FAP256"],"other_free_text":[]},"mechanistic_narrative":"MAK is a ciliary serine/threonine kinase of the CDC2-related family that functions as a master negative regulator of ciliary length by phosphorylating multiple substrates at the ciliary tip, including IFT-74 (reducing tubulin-binding affinity to promote tubulin unloading), RP1 (limiting axonemal extension), and FAP256/CEP104 (regulating axonemal microtubule assembly) [PMID:35969738, PMID:21148103, PMID:41231942]. MAK is activated through phosphorylation of its TDY activation-loop threonine by the upstream kinase CCRK, and functions cooperatively with ICK to maintain ciliary axoneme integrity in photoreceptors [PMID:16954377, PMID:39293864]. Loss of MAK causes elongated cilia with accumulated intraflagellar transport proteins, progressive photoreceptor degeneration, and retinitis pigmentosa in humans, with disease-causing mutations abolishing kinase activity [PMID:21148103, PMID:21835304, PMID:21825139]. Gene delivery of MAK or the functionally redundant ICK rescues ciliary length defects and retinal degeneration in patient-derived cells and animal models [PMID:34518651, PMID:39293864]."},"prefetch_data":{"uniprot":{"accession":"P20794","full_name":"Serine/threonine-protein kinase MAK","aliases":["Male germ cell-associated kinase"],"length_aa":623,"mass_kda":70.6,"function":"Essential for the regulation of ciliary length and required for the long-term survival of photoreceptors (By similarity). Phosphorylates FZR1 in a cell cycle-dependent manner. Plays a role in the transcriptional coactivation of AR. Could play an important function in spermatogenesis. May play a role in chromosomal stability in prostate cancer cells","subcellular_location":"Nucleus; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome; Cytoplasm, cytoskeleton, spindle; Midbody; Cell projection, cilium, photoreceptor outer segment; Photoreceptor inner segment","url":"https://www.uniprot.org/uniprotkb/P20794/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MAK","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MAK","total_profiled":1310},"omim":[{"mim_id":"620969","title":"ANEMIA, CONGENITAL DYSERYTHROPOIETIC, TYPE IVb; CDAN4B","url":"https://www.omim.org/entry/620969"},{"mim_id":"620825","title":"IMMUNODEFICIENCY 119; IMD119","url":"https://www.omim.org/entry/620825"},{"mim_id":"620271","title":"TRANSMEMBRANE PROTEIN 41B; TMEM41B","url":"https://www.omim.org/entry/620271"},{"mim_id":"619823","title":"ACIDIC NUCLEAR PHOSPHOPROTEIN 32 FAMILY, MEMBER B; ANP32B","url":"https://www.omim.org/entry/619823"},{"mim_id":"618774","title":"CEBALID SYNDROME; CEBALID","url":"https://www.omim.org/entry/618774"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Basal body","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"},{"location":"Connecting piece","reliability":"Approved"},{"location":"Nucleoli","reliability":"Additional"},{"location":"Plasma membrane","reliability":"Additional"},{"location":"Principal piece","reliability":"Additional"},{"location":"End piece","reliability":"Additional"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"retina","ntpm":110.1}],"url":"https://www.proteinatlas.org/search/MAK"},"hgnc":{"alias_symbol":["dJ417M14.2","RP62"],"prev_symbol":[]},"alphafold":{"accession":"P20794","domains":[{"cath_id":"3.30.200.20","chopping":"3-81","consensus_level":"high","plddt":89.1409,"start":3,"end":81},{"cath_id":"1.10.510.10","chopping":"84-289","consensus_level":"high","plddt":90.412,"start":84,"end":289}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P20794","model_url":"https://alphafold.ebi.ac.uk/files/AF-P20794-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P20794-F1-predicted_aligned_error_v6.png","plddt_mean":62.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MAK","jax_strain_url":"https://www.jax.org/strain/search?query=MAK"},"sequence":{"accession":"P20794","fasta_url":"https://rest.uniprot.org/uniprotkb/P20794.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P20794/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P20794"}},"corpus_meta":[{"pmid":"21825139","id":"PMC_21825139","title":"Exome sequencing and analysis of induced pluripotent stem cells identify the cilia-related gene male germ cell-associated kinase (MAK) as a cause of retinitis pigmentosa.","date":"2011","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/21825139","citation_count":177,"is_preprint":false},{"pmid":"21148103","id":"PMC_21148103","title":"Negative regulation of ciliary length by ciliary male germ cell-associated kinase (Mak) is required for retinal photoreceptor survival.","date":"2010","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/21148103","citation_count":117,"is_preprint":false},{"pmid":"2183027","id":"PMC_2183027","title":"A novel mammalian protein kinase gene (mak) is highly expressed in testicular germ cells at and after meiosis.","date":"1990","source":"Molecular and cellular 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pigmentosa caused by mutations in the ciliary MAK gene is relatively mild and is not associated with apparent extra-ocular features.","date":"2014","source":"Acta ophthalmologica","url":"https://pubmed.ncbi.nlm.nih.gov/25385675","citation_count":16,"is_preprint":false},{"pmid":"11129044","id":"PMC_11129044","title":"The MAK-V protein kinase regulates endocytosis in mouse.","date":"2000","source":"Molecular & general genetics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/11129044","citation_count":15,"is_preprint":false},{"pmid":"26558903","id":"PMC_26558903","title":"Efficient In Silico Identification of a Common Insertion in the MAK Gene which Causes Retinitis Pigmentosa.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26558903","citation_count":12,"is_preprint":false},{"pmid":"27268441","id":"PMC_27268441","title":"Involvement of MAK-1 and MAK-2 MAP kinases in cell wall integrity in Neurospora crassa.","date":"2016","source":"Bioscience, biotechnology, and 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Correlation of survival with MAK cell tumor necrosis factor production in vitro.","date":"1991","source":"Lymphokine and cytokine research","url":"https://pubmed.ncbi.nlm.nih.gov/1873360","citation_count":5,"is_preprint":false},{"pmid":"35253837","id":"PMC_35253837","title":"PCARE requires coiled coil, RP62 kinase-binding and EVH1 domain-binding motifs for ciliary expansion.","date":"2022","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/35253837","citation_count":4,"is_preprint":false},{"pmid":"31608171","id":"PMC_31608171","title":"Plastome of mycoheterotrophic Burmannia itoana Mak. (Burmanniaceae) exhibits extensive degradation and distinct rearrangements.","date":"2019","source":"PeerJ","url":"https://pubmed.ncbi.nlm.nih.gov/31608171","citation_count":4,"is_preprint":false},{"pmid":"21568852","id":"PMC_21568852","title":"Interaction between MAK-V protein kinase and synaptopodin.","date":"2011","source":"Biochemistry. 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Cancer & chemotherapy","url":"https://pubmed.ncbi.nlm.nih.gov/29066688","citation_count":1,"is_preprint":false},{"pmid":"41971568","id":"PMC_41971568","title":"Zinc diamyldithiocarbamate: MAK Value Documentation - Translation of the German version from 2023.","date":"2025","source":"The MAK collection for occupational health and safety","url":"https://pubmed.ncbi.nlm.nih.gov/41971568","citation_count":0,"is_preprint":false},{"pmid":"41584501","id":"PMC_41584501","title":"Tetramethylol acetylenediurea: MAK Value Documentation - Translation of the German version from 2023.","date":"2025","source":"The MAK collection for occupational health and safety","url":"https://pubmed.ncbi.nlm.nih.gov/41584501","citation_count":0,"is_preprint":false},{"pmid":"41231942","id":"PMC_41231942","title":"Chlamydomonas protein kinase MAK phosphorylates FAP256/CEP104 and regulates axonemal microtubule assembly.","date":"2025","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/41231942","citation_count":0,"is_preprint":false},{"pmid":"15773550","id":"PMC_15773550","title":"[Molecular cloning and characterization of the mouse mak-v/Hunk gene promoter].","date":"2005","source":"Molekuliarnaia biologiia","url":"https://pubmed.ncbi.nlm.nih.gov/15773550","citation_count":0,"is_preprint":false},{"pmid":"41971562","id":"PMC_41971562","title":"Synthetic amorphous silica: MAK Value Documentation, addendum - Translation of the German version from 2023.","date":"2025","source":"The MAK collection for occupational health and safety","url":"https://pubmed.ncbi.nlm.nih.gov/41971562","citation_count":0,"is_preprint":false},{"pmid":"41586081","id":"PMC_41586081","title":"N,N-Dimethyl-p-toluidine: MAK Value Documentation - Translation of the German version from 2022.","date":"2025","source":"The MAK collection for occupational health and safety","url":"https://pubmed.ncbi.nlm.nih.gov/41586081","citation_count":0,"is_preprint":false},{"pmid":"41575991","id":"PMC_41575991","title":"Formononetin ameliorates SP-induced urticaria in mice via suppressing TAK1/MAK signaling pathway.","date":"2026","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/41575991","citation_count":0,"is_preprint":false},{"pmid":"12068635","id":"PMC_12068635","title":"[The use of the two-hybrid cloning in yeast for functional characterization of protein kinase MAK-V].","date":"2002","source":"Molekuliarnaia biologiia","url":"https://pubmed.ncbi.nlm.nih.gov/12068635","citation_count":0,"is_preprint":false},{"pmid":"8679938","id":"PMC_8679938","title":"Protoplast culture and plant regeneration from the suspension cells of Gynostemma pentaphyllum (Thumb) Mak.","date":"1995","source":"Chinese journal of biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/8679938","citation_count":0,"is_preprint":false},{"pmid":"125971","id":"PMC_125971","title":"[Characterisation of RNA from synchronously growing yeast cells (Saccharomyces cerevisiae) as separated by columns of methylated albumin on Kieselgur (MAK) (author's transl)].","date":"1975","source":"Zeitschrift fur Naturforschung. Section C, Biosciences","url":"https://pubmed.ncbi.nlm.nih.gov/125971","citation_count":0,"is_preprint":false},{"pmid":"41973600","id":"PMC_41973600","title":"Chloroform: MAK Value Documentation, addendum - Translation of the German version from 2022.","date":"2025","source":"The MAK collection for occupational health and safety","url":"https://pubmed.ncbi.nlm.nih.gov/41973600","citation_count":0,"is_preprint":false},{"pmid":"41657991","id":"PMC_41657991","title":"Single-cell RNA-seq reveals a key role for Vibrio cholerae Mak toxins in Tetrahymena pyriformis killing and bacterial survival.","date":"2026","source":"Frontiers in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/41657991","citation_count":0,"is_preprint":false},{"pmid":"41586701","id":"PMC_41586701","title":"Naphthalene: MAK Value Documentation, addendum - Translation of the German version from 2022.","date":"2025","source":"The MAK collection for occupational health and safety","url":"https://pubmed.ncbi.nlm.nih.gov/41586701","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.10.08.681228","title":"DeepEmbCas9: Cas9 coevolution and sgRNA structural information for CRISPR-Cas9 cleavage activity prediction","date":"2025-10-08","source":"bioRxiv","url":"https://doi.org/10.1101/2025.10.08.681228","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.11.20.689482","title":"Metabolic engineering of  <i>Corynebacterium glutamicum</i>  for production of the low-caloric natural sweetener D-allulose via phosphorylated intermediates","date":"2025-11-20","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.20.689482","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":38580,"output_tokens":3401,"usd":0.083377},"stage2":{"model":"claude-opus-4-6","input_tokens":6764,"output_tokens":2930,"usd":0.160605},"total_usd":0.243982,"stage1_batch_id":"msgbatch_01Mgbec4vM6c9vXht7JVJzss","stage2_batch_id":"msgbatch_01JkuuH5RB2BdJNsgnhjbC6s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1990,\n      \"finding\": \"MAK (male germ cell-associated kinase) encodes a novel serine/threonine protein kinase whose kinase domain shares ~40% identity with the cdc2-CDC28 gene family; its transcripts are expressed almost exclusively in testicular germ cells entering and after meiosis.\",\n      \"method\": \"cDNA cloning, sequence analysis, Northern blot\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — original cloning with sequence analysis establishing kinase identity and expression pattern, foundational paper with 113 citations\",\n      \"pmids\": [\"2183027\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"MAK protein products (66 and 60 kDa) are most abundant in late pachytene spermatocytes, localize as soluble cytoplasmic proteins, associate with a 210 kDa cellular protein, and phosphorylate this 210 kDa protein on serine and threonine residues both in vitro (immunoprecipitate kinase assay) and in vivo (32Pi labeling).\",\n      \"method\": \"Subcellular fractionation, immunoprecipitation kinase assay, [35S]methionine labeling, 32Pi metabolic labeling\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro kinase assay combined with in vivo phosphorylation and fractionation, replicated with multiple antibodies\",\n      \"pmids\": [\"8321219\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"MAK and MRK/ICK require dual phosphorylation in a TDY activation-loop motif: threonine phosphorylation (T157 in MRK) is catalyzed by CCRK (cell cycle-related kinase), while tyrosine phosphorylation is by autophosphorylation. Protein phosphatase 5 (PP5) dephosphorylates MRK at T157, acting as the opposing regulator. MRK preferentially phosphorylates R-P-X-S/T-P motifs (consensus determined by combinatorial peptide library screening), and phosphorylates the antiapoptotic protein Scythe at T1080.\",\n      \"method\": \"In vitro kinase assay, combinatorial peptide library screening, site-directed mutagenesis, mass spectrometry, co-immunoprecipitation\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstituted kinase cascade in vitro with mutagenesis and MS validation, multiple orthogonal methods in single study\",\n      \"pmids\": [\"16954377\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Mak localizes to the connecting cilia and outer-segment axonemes of photoreceptor cells and negatively regulates ciliary length. In Mak-null retinas, photoreceptors show elongated cilia with accumulated IFT88, IFT57, expanded Kif3a, and acetylated α-tubulin signals, and rhodopsin accumulates abnormally in cell bodies. Overexpression of RP1 (a microtubule-associated protein) induces ciliary elongation, and Mak coexpression rescues this; the RP1 N-terminal portion is phosphorylated by Mak in vitro, linking Mak kinase activity to ciliary length control.\",\n      \"method\": \"Mak-null mouse model, immunohistochemistry, in vitro kinase assay, overexpression rescue experiments\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — genetic KO with defined cellular phenotype, in vitro kinase assay identifying RP1 as substrate, rescue experiments; 117 citations\",\n      \"pmids\": [\"21148103\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Missense mutations in MAK that alter conserved kinase residues abolish kinase activity as demonstrated by in vitro kinase assay, establishing that MAK kinase activity is essential for its function in photoreceptor ciliary maintenance.\",\n      \"method\": \"In vitro kinase assay with disease-causing mutants\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro kinase assay directly linking catalytic activity to human disease mutations\",\n      \"pmids\": [\"21835304\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"MAK is expressed in the inner segments, cell bodies, and axons of rod and cone photoreceptors in human donor tissue. An exon 9-containing MAK isoform (also including a novel exon 12) is the predominant transcript in retinal precursor cells, and failure to switch to this isoform (caused by an Alu insertion in exon 9) results in loss of normal MAK protein and retinitis pigmentosa.\",\n      \"method\": \"Immunohistochemistry of human donor tissue, iPSC differentiation into retinal precursors, RT-PCR isoform analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct localization in human tissue plus functional isoform analysis in iPSC-derived retinal precursors; 177 citations\",\n      \"pmids\": [\"21825139\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"MAK-deficient (Mak-/-) mice are viable and fertile with intact spermatogenesis, demonstrating that MAK is not essential for sperm formation, though Mak-/- males show mildly reduced litter sizes and reduced sperm motility in vitro.\",\n      \"method\": \"Targeted gene knockout, fertility and spermatogenesis phenotyping\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean genetic KO with defined phenotypic readouts in multiple assays\",\n      \"pmids\": [\"11971961\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In C. elegans, DYF-18/CCRK and DYF-5/MAK act in a kinase cascade to regulate ciliary length and branching morphology: loss of dyf-18 or dyf-5 results in dramatically elongated, unbranched AWA olfactory neuron cilia. The mechanism involves regulation of axonemal microtubule stability, as EBP-2 end-binding protein decorates axonemal microtubules along their lengths, tubulin load increases, and tubulin turnover is reduced in dyf-18 mutants. Microtubule-destabilizing tubulin mutations and IFT protein mutations that disrupt tubulin transport suppress AWA cilia elongation in dyf-18 mutants.\",\n      \"method\": \"C. elegans genetic epistasis, live imaging, IFT motor localization, genetic suppressor analysis\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — genetic epistasis with multiple suppressor mutations plus live imaging defining mechanism\",\n      \"pmids\": [\"30955935\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The ciliary kinase DYF-5/MAK phosphorylates multiple sites within the tubulin-binding module of IFT-74, reducing its tubulin-binding affinity approximately sixfold and promoting tubulin unloading at the ciliary tip. Ablation or constitutive activation of IFT-74 phosphorylation abnormally elongates or shortens sensory cilia in C. elegans neurons, respectively.\",\n      \"method\": \"In vitro phosphorylation assay, affinity binding measurements, C. elegans genetic analysis with phosphomimetic/phosphoablating IFT-74 mutations\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution of phosphorylation event with quantified affinity change, validated in vivo with gain/loss-of-function mutations\",\n      \"pmids\": [\"35969738\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MAK localizes to the ciliary tip and functions as an IFT regulator cooperatively with ICK. Simultaneous disruption of Mak and Ick in mice results in loss of photoreceptor ciliary axonemes and severe retinal degeneration. CCRK kinase is identified as an upstream activator of both Mak and Ick in retinal photoreceptor cells. Gene delivery of Ick and pharmacological inhibition of FGFR (a negative regulator of Ick) ameliorate retinal degeneration in Mak-/- mice.\",\n      \"method\": \"Double-knockout mouse model, live imaging, subretinal gene delivery, pharmacological FGFR inhibition\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic double-KO with defined phenotype, pathway placement via epistasis, therapeutic rescue validates mechanism\",\n      \"pmids\": [\"39293864\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Viral overexpression of the retinal MAK transgene in patient iPSC-derived photoreceptor precursor cells restores MAK transcript and protein. Overexpression of retinal MAK in patient-derived fibroblasts and mak-knockdown zebrafish restores the ability to regulate primary cilia length, and mitigates the visual defect in mak knockdown zebrafish.\",\n      \"method\": \"iPSC-derived cells, viral gene delivery, zebrafish morpholino knockdown, cilia length assay\",\n      \"journal\": \"Gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — functional rescue of cilia length defect in patient cells and zebrafish model, but single lab study\",\n      \"pmids\": [\"34518651\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PCARE contains evolutionary conserved binding sites for the photoreceptor kinase MAK at its C-terminal region. Deletion of the MAK-binding motifs in PCARE did not abolish ciliary localization but affected downstream WASF3-mediated ciliary tip membrane expansion, suggesting MAK interacts with PCARE to regulate actin-driven photoreceptor outer segment disk formation.\",\n      \"method\": \"Co-expression studies, deletion mutagenesis, ciliary expansion assay\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — functional co-expression with deletion mutagenesis, but direct binding not biochemically confirmed with pulldown\",\n      \"pmids\": [\"35253837\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In Chlamydomonas, CrMAK (ortholog of mammalian ICK/MAK) localizes to cilia and is enriched at the ciliary tip during active ciliary assembly. CrMAK associates with the axoneme via its C-terminal tail, acts downstream of the LF1/LF2/LF3 ciliary length-regulatory complex, and phosphorylates FAP256/CEP104 (a microtubule plus-end tracking protein). Loss of CrMAK or its kinase activity leads to aciliated cells, and dephosphorylation of FAP256/CEP104 impairs ciliary assembly.\",\n      \"method\": \"Chlamydomonas mutant analysis, in vitro kinase assay, co-immunoprecipitation, live imaging, mutagenesis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution of phosphorylation with identified substrate (FAP256/CEP104), genetic epistasis placing MAK downstream of LF complex, loss-of-function phenotype\",\n      \"pmids\": [\"41231942\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MAK (male germ cell-associated kinase) is a serine/threonine kinase activated by CCRK-mediated phosphorylation of its TDY activation-loop threonine, that localizes to the ciliary tip where it phosphorylates IFT-74 (reducing tubulin-binding affinity to promote tubulin unloading), RP1/microtubule-associated proteins (to limit axonemal extension), and FAP256/CEP104 (to regulate axonemal microtubule assembly), thereby acting as a master negative regulator of ciliary length whose loss causes progressive photoreceptor degeneration and retinitis pigmentosa.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MAK is a ciliary serine/threonine kinase of the CDC2-related family that functions as a master negative regulator of ciliary length by phosphorylating multiple substrates at the ciliary tip, including IFT-74 (reducing tubulin-binding affinity to promote tubulin unloading), RP1 (limiting axonemal extension), and FAP256/CEP104 (regulating axonemal microtubule assembly) [PMID:35969738, PMID:21148103, PMID:41231942]. MAK is activated through phosphorylation of its TDY activation-loop threonine by the upstream kinase CCRK, and functions cooperatively with ICK to maintain ciliary axoneme integrity in photoreceptors [PMID:16954377, PMID:39293864]. Loss of MAK causes elongated cilia with accumulated intraflagellar transport proteins, progressive photoreceptor degeneration, and retinitis pigmentosa in humans, with disease-causing mutations abolishing kinase activity [PMID:21148103, PMID:21835304, PMID:21825139]. Gene delivery of MAK or the functionally redundant ICK rescues ciliary length defects and retinal degeneration in patient-derived cells and animal models [PMID:34518651, PMID:39293864].\",\n  \"teleology\": [\n    {\n      \"year\": 1990,\n      \"claim\": \"Identification of MAK as a novel CDC2-related serine/threonine kinase expressed predominantly in meiotic germ cells established the gene's kinase identity and pointed to a specialized role in differentiated cell types.\",\n      \"evidence\": \"cDNA cloning, sequence analysis, and Northern blot in mouse testis\",\n      \"pmids\": [\"2183027\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No substrate or pathway identified\", \"Function in non-testicular tissues unknown\"]\n    },\n    {\n      \"year\": 1993,\n      \"claim\": \"Demonstration that MAK associates with and phosphorylates a 210 kDa protein in spermatocytes both in vitro and in vivo established that MAK functions as an active kinase with at least one endogenous substrate in germ cells.\",\n      \"evidence\": \"Immunoprecipitate kinase assay and 32Pi metabolic labeling in pachytene spermatocytes\",\n      \"pmids\": [\"8321219\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the 210 kDa substrate unknown\", \"Physiological role of this phosphorylation unclear\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"The finding that Mak-knockout mice are viable and fertile with only mild sperm motility defects redirected the field away from an essential spermatogenesis role and opened the question of MAK's primary physiological function.\",\n      \"evidence\": \"Targeted gene knockout in mice with fertility and spermatogenesis phenotyping\",\n      \"pmids\": [\"11971961\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Non-testicular phenotypes not yet examined\", \"Functional redundancy with ICK not tested\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Reconstitution of the CCRK→MAK/MRK activation cascade, including identification of the TDY activation-loop mechanism, PP5 as opposing phosphatase, and the R-P-X-S/T-P substrate consensus, defined how MAK is activated and what it recognizes.\",\n      \"evidence\": \"In vitro kinase assay, combinatorial peptide library screening, mutagenesis, mass spectrometry\",\n      \"pmids\": [\"16954377\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological substrates matching R-P-X-S/T-P consensus not identified in vivo\", \"Whether CCRK activates MAK in all tissues unknown\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Discovery that Mak localizes to photoreceptor connecting cilia and negatively regulates ciliary length — with Mak-null retinas showing elongated cilia, IFT accumulation, and rhodopsin mislocalization — fundamentally recast MAK as a ciliary kinase, with RP1 identified as a direct phospho-substrate linking kinase activity to axonemal length control.\",\n      \"evidence\": \"Mak-null mouse model, immunohistochemistry, in vitro kinase assay, overexpression rescue\",\n      \"pmids\": [\"21148103\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether RP1 phosphorylation alone accounts for length phenotype unknown\", \"Mechanism of IFT accumulation not defined\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Two studies linked MAK loss-of-function to human retinitis pigmentosa: kinase-dead missense mutations were shown to abolish catalytic activity, and an Alu insertion disrupting retinal MAK isoform splicing was identified as a disease-causing allele, establishing MAK as a retinitis pigmentosa gene requiring kinase activity for photoreceptor survival.\",\n      \"evidence\": \"In vitro kinase assay with patient mutations; iPSC-derived retinal precursor isoform analysis; immunohistochemistry of human donor retina\",\n      \"pmids\": [\"21835304\", \"21825139\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Natural history and genotype-phenotype correlations incompletely characterized\", \"Whether cone versus rod photoreceptors are differentially affected unclear\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Genetic epistasis in C. elegans demonstrated that the CCRK→MAK cascade controls ciliary length by regulating axonemal microtubule stability and tubulin delivery via IFT, with microtubule-destabilizing mutations and IFT mutations suppressing the elongation phenotype.\",\n      \"evidence\": \"C. elegans genetic epistasis, live imaging, suppressor analysis in AWA olfactory neurons\",\n      \"pmids\": [\"30955935\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct phosphorylation target mediating microtubule stability not identified in this study\", \"Whether branching versus length regulation are separable unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Viral delivery of retinal MAK rescued cilia length defects in patient iPSC-derived photoreceptor precursors and visual function in mak-knockdown zebrafish, providing proof-of-concept for gene therapy.\",\n      \"evidence\": \"iPSC-derived cells, viral gene delivery, zebrafish morpholino knockdown, cilia length assay\",\n      \"pmids\": [\"34518651\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Long-term efficacy and safety of gene delivery not assessed\", \"Single-lab study awaits independent replication\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identification of IFT-74 as a direct MAK/DYF-5 substrate whose phosphorylation reduces tubulin-binding affinity ~6-fold provided a molecular mechanism for how MAK promotes tubulin unloading at the ciliary tip to control axonemal length.\",\n      \"evidence\": \"In vitro phosphorylation assay, affinity measurements, phosphomimetic/phosphoablating IFT-74 mutations in C. elegans\",\n      \"pmids\": [\"35969738\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether IFT-74 phosphorylation operates identically in mammalian photoreceptors not shown\", \"Contribution of IFT-74 phosphorylation relative to RP1 phosphorylation unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"PCARE was identified as a MAK-interacting protein at the ciliary tip, with MAK-binding motifs required for WASF3-mediated membrane expansion, suggesting MAK regulates actin-driven photoreceptor outer segment disk formation.\",\n      \"evidence\": \"Co-expression studies, deletion mutagenesis, ciliary expansion assay\",\n      \"pmids\": [\"35253837\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical binding not confirmed by pulldown or co-IP\", \"Whether MAK phosphorylates PCARE not tested\", \"Single co-expression system without in vivo validation\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Double knockout of Mak and Ick in mice revealed functional cooperativity between these kinases, with combined loss causing complete loss of photoreceptor ciliary axonemes and severe degeneration; Ick gene delivery and pharmacological FGFR inhibition rescued Mak-null retinal degeneration, establishing therapeutic redundancy.\",\n      \"evidence\": \"Double-knockout mouse model, subretinal gene delivery, pharmacological FGFR inhibition\",\n      \"pmids\": [\"39293864\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether MAK and ICK share identical substrates or have distinct targets not resolved\", \"Optimal therapeutic window for intervention not defined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"In Chlamydomonas, CrMAK was placed downstream of the LF1/LF2/LF3 length-regulatory complex and shown to phosphorylate FAP256/CEP104, a microtubule plus-end tracking protein, whose dephosphorylation impairs ciliary assembly — adding a third identified substrate to MAK's ciliary repertoire.\",\n      \"evidence\": \"Chlamydomonas mutant analysis, in vitro kinase assay, co-immunoprecipitation, live imaging\",\n      \"pmids\": [\"41231942\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether mammalian CEP104 is similarly regulated by MAK not tested\", \"Structural basis for CrMAK axonemal association via C-terminal tail unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The relative contributions of MAK's multiple substrates (IFT-74, RP1, FAP256/CEP104, PCARE) to ciliary length control and photoreceptor outer segment homeostasis remain unintegrated, and no structural model of MAK exists to explain substrate selectivity or the cooperative relationship with ICK.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No crystal or cryo-EM structure of MAK\", \"Quantitative contribution of each substrate to ciliary length phenotype not parsed\", \"Whether MAK has non-ciliary functions in retinal or other neurons remains open\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 2, 3, 4, 8, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [3, 5, 9, 12]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [3, 7, 8, 9, 12]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [3, 7, 8, 9, 12]},\n      {\"term_id\": \"R-HSA-9709957\", \"supporting_discovery_ids\": [3, 5, 9, 10]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"CCRK\", \"ICK\", \"RP1\", \"IFT74\", \"PCARE\", \"FAP256\"],\n    \"other_free_text\": []\n  }\n}\n```"}