Affinage

DNAAF5

Dynein axonemal assembly factor 5 · UniProt Q86Y56

Round 2 corrected
Length
855 aa
Mass
93.5 kDa
Annotated
2026-04-28
38 papers in source corpus 4 papers cited in narrative 4 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

DNAAF5 (formerly HEATR2) is a cytoplasmic dynein axonemal assembly factor required for the pre-assembly and/or transport of dynein arms into motile cilia; its loss abolishes outer dynein arms and ciliary beating, causing primary ciliary dyskinesia (PMID:23040496). CRISPR knock-in mouse models demonstrate allele-specific and tissue-specific requirements: null alleles are embryonic lethal, whereas hypomorphic missense alleles permit partial cilia function, and quantitative proteomics of mutant airway cilia reveal broader reductions in axonemal regulatory and structural proteins beyond dynein arms (PMID:37104040). In hepatocellular carcinoma cells, DNAAF5 additionally functions as a scaffold that recruits the deubiquitinase USP39 to stabilize the glycolytic enzyme PFKL, promoting tumor cell proliferation and sorafenib resistance (PMID:36276075).

Mechanistic history

Synthesis pass · year-by-year structured walk · 4 steps
  1. 2012 High

    The fundamental question of DNAAF5's cellular role was answered: it is a cytoplasmic factor required for outer dynein arm assembly in motile cilia, establishing that ciliary dynein arm biogenesis depends on dedicated cytoplasmic assembly factors beyond known axonemal structural components.

    Evidence Whole-exome sequencing of PCD patients, miRNA knockdown in Chlamydomonas, shRNA knockdown in human airway epithelia, immunohistochemistry, and TEM across human and algal systems

    PMID:23040496

    Open questions at the time
    • Precise molecular mechanism of DNAAF5 in dynein pre-assembly (direct chaperone vs. scaffold vs. transport adaptor) was not resolved
    • Whether DNAAF5 also contributes to inner dynein arm assembly was not definitively tested
    • No structural information on DNAAF5 or its interaction partners in the dynein assembly pathway
  2. 2022 Medium

    A non-ciliary scaffolding function for DNAAF5 was identified in hepatocellular carcinoma, showing it recruits USP39 to deubiquitinate and stabilize PFKL, linking DNAAF5 to glycolytic regulation and tumor proliferation — a role mechanistically distinct from its ciliary assembly function.

    Evidence Co-immunoprecipitation, mass spectrometry, overexpression/knockout cell lines, colony formation assays, and xenograft models in hepatocellular carcinoma

    PMID:36276075

    Open questions at the time
    • Single-lab study; DNAAF5–USP39–PFKL ternary complex not independently replicated
    • Whether the DNAAF5 scaffolding role for USP39 operates in non-cancerous tissues is unknown
    • Structural basis for DNAAF5 interaction with both USP39 and PFKL not determined
  3. 2023 High

    Allele-specific and tissue-specific requirements for DNAAF5 were established, revealing that complete loss is embryonic lethal while hypomorphic alleles produce graded ciliary dysfunction, and that DNAAF5 function extends to axonemal regulatory and structural proteins beyond dynein arms.

    Evidence CRISPR-Cas9 knock-in mouse models (null and missense alleles), quantitative proteomics of isolated airway cilia, transcriptomic profiling of mouse and human mutant cells

    PMID:37104040

    Open questions at the time
    • Whether the broader proteomic changes reflect a direct DNAAF5 function in non-dynein assembly or are secondary to dynein loss is unresolved
    • Compensatory transcriptional upregulation mechanisms are not molecularly defined
    • Genotype–phenotype relationship for diverse human DNAAF5 alleles remains incompletely mapped
  4. 2025 Medium

    Conserved expression of dnaaf5 in motile ciliated tissues and ciliopathic phenotypes in zebrafish crispants confirmed that DNAAF5's role in motile cilia biogenesis is broadly conserved across vertebrates.

    Evidence Whole-mount in situ hybridization and CRISPR crispant phenotyping in zebrafish

    PMID:42029186

    Open questions at the time
    • Zebrafish study is from a single lab; independent replication pending
    • Molecular interactors of dnaaf5 in zebrafish not identified
    • Relationship between zebrafish dnaaf5 crispant phenotypes and specific dynein arm subtypes not resolved

Open questions

Synthesis pass · forward-looking unresolved questions
  • The direct molecular mechanism by which DNAAF5 participates in dynein arm pre-assembly — whether as a chaperone, scaffold for a multi-protein assembly complex, or transport adaptor — remains undefined, and no structural model of DNAAF5 or its complexes exists.
  • No crystal or cryo-EM structure of DNAAF5 alone or in complex
  • Identity of the full set of direct binding partners in the dynein assembly pathway is incomplete
  • Relationship between the ciliary assembly function and the oncogenic scaffolding function has not been investigated

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 2
Localization
GO:0005829 cytosol 1
Pathway
R-HSA-1852241 Organelle biogenesis and maintenance 3 R-HSA-1643685 Disease 1
Partners
PFKLUSP39

Evidence

Reading pass · 4 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2012 HEATR2 (DNAAF5) was identified as a dynein axonemal assembly factor required for outer dynein arm (ODA) assembly in motile cilia. Loss-of-function in patients and model organisms resulted in absent dynein arms and loss of ciliary beating. Immunohistochemistry localized HEATR2 to the cytoplasm (not within cilia), indicating a role in cytoplasmic pre-assembly or transport of dynein arms rather than as a structural axonemal component. MicroRNA-mediated silencing of the Chlamydomonas ortholog and shRNA knockdown in human airway epithelial cells recapitulated absent ODAs and reduced flagellar beat frequency. Whole-exome sequencing of PCD patients, immunohistochemistry for subcellular localization, miRNA knockdown in Chlamydomonas reinhardtii, shRNA knockdown in human airway epithelial cells, transmission electron microscopy of axoneme ultrastructure, ciliary beat frequency measurement American journal of human genetics High 23040496
2023 Allele-specific and tissue-specific requirements for DNAAF5 in dynein motor assembly were established using CRISPR-Cas9 mouse models recreating a human missense variant and a frameshift-null allele. Homozygous null Dnaaf5 was embryonic lethal; compound heterozygous (missense/null) animals showed severe hydrocephalus and early death; homozygous missense animals had partial cilia function and improved survival. Proteomic analysis of airway cilia from mutant mice revealed reductions in axonemal regulatory and structural proteins not previously linked to DNAAF5, and transcriptional analysis showed compensatory upregulation of axonemal protein-coding genes in mutant cells. CRISPR-Cas9 knock-in mouse models, ultrastructure analysis (electron microscopy), quantitative proteomics of isolated airway cilia, transcriptional (RNA) analysis of mouse and human mutant cells JCI insight High 37104040
2022 In hepatocellular carcinoma cells, DNAAF5 was found to act as a scaffold protein that directly binds PFKL (phosphofructokinase, liver type) and recruits the deubiquitinase USP39, forming a ternary complex. This interaction stabilizes PFKL protein by promoting its deubiquitination, thereby enhancing glycolytic activity and tumor cell proliferation. DNAAF5 knockout reduced proliferation and increased sorafenib sensitivity, and USP39 knockdown suppressed the proliferative effect of DNAAF5 overexpression in vitro and in vivo. Co-immunoprecipitation, mass spectrometry (transcriptome sequencing + proteomics), stable cell line overexpression/knockout, colony formation assay, xenograft in vivo model, tissue microarray Frontiers in oncology Medium 36276075
2025 Zebrafish dnaaf5 mRNA is expressed in motile ciliated tissues including Kupffer's vesicle, pronephros, floor plate, brain, and olfactory placode. CRISPR-based crispants for dnaaf5 developed ciliopathic defects, consistent with a conserved role in motile cilia biogenesis and function in vertebrates. Whole-mount in situ hybridization for spatio-temporal expression mapping, CRISPR crispant generation, phenotypic analysis of ciliopathic defects in zebrafish embryos The International journal of developmental biology Medium 42029186

Source papers

Stage 0 corpus · 38 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2012 Insights into RNA biology from an atlas of mammalian mRNA-binding proteins. Cell 1718 22658674
2005 A human protein-protein interaction network: a resource for annotating the proteome. Cell 1704 16169070
2002 Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. Proceedings of the National Academy of Sciences of the United States of America 1479 12477932
2015 The BioPlex Network: A Systematic Exploration of the Human Interactome. Cell 1118 26186194
2017 Architecture of the human interactome defines protein communities and disease networks. Nature 1085 28514442
2015 A human interactome in three quantitative dimensions organized by stoichiometries and abundances. Cell 1015 26496610
2003 Complete sequencing and characterization of 21,243 full-length human cDNAs. Nature genetics 754 14702039
2007 Large-scale mapping of human protein-protein interactions by mass spectrometry. Molecular systems biology 733 17353931
2021 Dual proteome-scale networks reveal cell-specific remodeling of the human interactome. Cell 705 33961781
2011 Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium. Briefings in bioinformatics 656 21873635
1994 Oligo-capping: a simple method to replace the cap structure of eukaryotic mRNAs with oligoribonucleotides. Gene 492 8125298
2004 The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC). Genome research 438 15489334
2015 Panorama of ancient metazoan macromolecular complexes. Nature 407 26344197
2021 A proximity-dependent biotinylation map of a human cell. Nature 339 34079125
2017 Genome-wide CRISPR screen identifies HNRNPL as a prostate cancer dependency regulating RNA splicing. Proceedings of the National Academy of Sciences of the United States of America 282 28611215
2012 A high-throughput approach for measuring temporal changes in the interactome. Nature methods 273 22863883
2016 An organelle-specific protein landscape identifies novel diseases and molecular mechanisms. Nature communications 211 27173435
2020 UFMylation maintains tumour suppressor p53 stability by antagonizing its ubiquitination. Nature cell biology 168 32807901
2019 A protein-interaction network of interferon-stimulated genes extends the innate immune system landscape. Nature immunology 159 30833792
2008 Systematic identification of mRNAs recruited to argonaute 2 by specific microRNAs and corresponding changes in transcript abundance. PloS one 148 18461144
2020 Comparative Application of BioID and TurboID for Protein-Proximity Biotinylation. Cells 146 32344865
2012 Whole-exome capture and sequencing identifies HEATR2 mutation as a cause of primary ciliary dyskinesia. American journal of human genetics 138 23040496
2021 A protein interaction landscape of breast cancer. Science (New York, N.Y.) 111 34591612
2021 Systematically defining selective autophagy receptor-specific cargo using autophagosome content profiling. Molecular cell 105 33545068
2020 The Hsp70-Hsp90 co-chaperone Hop/Stip1 shifts the proteostatic balance from folding towards degradation. Nature communications 101 33239621
2014 Proteomic analysis of the epidermal growth factor receptor (EGFR) interactome and post-translational modifications associated with receptor endocytosis in response to EGF and stress. Molecular & cellular proteomics : MCP 99 24797263
2021 SARS-CoV-2-host proteome interactions for antiviral drug discovery. Molecular systems biology 86 34709727
2017 A Genome-Wide CRISPR Screen Identifies Genes Critical for Resistance to FLT3 Inhibitor AC220. Cancer research 76 28625976
2015 Temporal proteomics of NGF-TrkA signaling identifies an inhibitory role for the E3 ligase Cbl-b in neuroblastoma cell differentiation. Science signaling 61 25921289
2020 Extensive rewiring of the EGFR network in colorectal cancer cells expressing transforming levels of KRASG13D. Nature communications 60 31980649
2023 The effect of Dnaaf5 gene dosage on primary ciliary dyskinesia phenotypes. JCI insight 17 37104040
2022 DNAAF5 promotes hepatocellular carcinoma malignant progression by recruiting USP39 to improve PFKL protein stability. Frontiers in oncology 16 36276075
2023 Genome-Wide Meta-Analysis of Cerebrospinal Fluid Biomarkers in Alzheimer's Disease and Parkinson's Disease Cohorts. Movement disorders : official journal of the Movement Disorder Society 9 37539664
2024 Identification of candidate genes affecting the tibia quality in Nonghua duck. Poultry science 3 38350390
2023 Genome-wide meta-analysis of CSF biomarkers in Alzheimer's disease and Parkinson's disease cohorts. medRxiv : the preprint server for health sciences 1 37398091
2025 Comprehensive Proteomics and Machine Learning Analysis to Distinguish Follicular Adenoma and Follicular Thyroid Carcinoma from Indeterminate Thyroid Nodules. Endocrinology and metabolism (Seoul, Korea) 0 40205804
2025 Dynein axonemal assembly factors (dnaaf) 5 and 9 are expressed in ciliated organs of zebrafish embryos. The International journal of developmental biology 0 42029186
2023 The effect of Dnaaf5 gene dosage on primary ciliary dyskinesia phenotypes. bioRxiv : the preprint server for biology 0 36712068