Affinage

CDC45

Cell division control protein 45 homolog · UniProt O75419

Length
566 aa
Mass
65.6 kDa
Annotated
2026-06-09
100 papers in source corpus 54 papers cited in narrative 54 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CDC45 is an essential eukaryotic DNA replication factor that, together with MCM2-7 and GINS, constitutes the CMG (Cdc45/Mcm2-7/GINS) replicative helicase (PMID:16798881, PMID:20122406). CDC45 and GINS bridge the Mcm2/Mcm5 gate of the helicase ring to form a topologically closed assembly, and this association elevates MCM2-7 ATP hydrolysis ~100-fold and converts the ring into a processive 3'-to-5' helicase that translocates on the leading-strand template (PMID:20122406, PMID:21378962, PMID:22474384). CDC45 retains a RecJ-like (DHH-family) fold whose ssDNA-binding groove guards the leading strand within the CMG, and mutations in this fold diminish helicase activity (PMID:27189187, PMID:25561522, PMID:22147708). CDC45 is loaded at origins in a stepwise, DDK- and CDK-dependent manner downstream of pre-RC assembly, requiring Mcm10, Sld3, TopBP1/Dpb11, and DONSON, with GINS displacing Sld3/Dpb11 to mature the CMS intermediate into the active CMG (PMID:11296242, PMID:11864598, PMID:16990792, PMID:21362622, PMID:37638758, PMID:25659432); the timing of CDC45 origin association is the key determinant of origin firing time and is rate-limiting for initiation (PMID:22169533, PMID:26919204). Beyond unwinding, CDC45 coordinates polymerase recruitment, tethering DNA polymerase alpha onto MCM via direct binding, supporting sequential loading of RPA, pol alpha, and PCNA, and catalytically loading RPA onto nascent ssDNA (PMID:9755170, PMID:10886370, PMID:28100698, PMID:10518787). CDC45 travels with the replisome throughout elongation and acts as a checkpoint hub: Chk2 phosphorylates CMG subunits to inhibit helicase activity, Chk1 displaces CDC45 from chromatin, and the CDC45 disordered loop recruits Rad53/checkpoint kinase to stalled forks (PMID:15329670, PMID:22853956, PMID:16912045, PMID:30595439). Biallelic partial loss-of-function mutations in CDC45 cause Meier-Gorlin syndrome and craniosynostosis, and CDC45 is essential for mammalian post-implantation development (PMID:27374770, PMID:11416137).

Mechanistic history

Synthesis pass · year-by-year structured walk · 12 steps
  1. 1997 High

    Established CDC45 as a genetically required initiation factor acting after START and downstream of pre-RC components, answering where in the cell cycle CDC45 functions.

    Evidence Yeast genetics, synthetic lethality with ORC/MCM alleles, 2D gel origin firing analysis, and CDC7/Dbf4 epistasis

    PMID:9001208 PMID:9356482

    Open questions at the time
    • Did not define a biochemical activity for CDC45
    • Mechanism of CDC45-dependent origin activation unresolved
  2. 2000 High

    Demonstrated that CDC45 drives the origin-unwinding step and orders the loading of downstream replication factors, distinguishing helicase activation from polymerase recruitment.

    Evidence Xenopus egg extract immunodepletion, supercoiling/unwinding assays, chromatin fractionation, and aphidicolin uncoupling

    PMID:10882098 PMID:10886370 PMID:9755170

    Open questions at the time
    • Did not identify the helicase enzyme itself
    • How CDC45 physically couples to MCM unclear at this stage
  3. 2006 High

    Identified CDC45-MCM2-7-GINS as a single stable complex (CMG) that is the eukaryotic replicative helicase, resolving the long-standing question of the replicative helicase identity.

    Evidence Immunoaffinity purification with MS, in vitro helicase assay on purified Drosophila CMG, and RNAi knockdown

    PMID:16483939 PMID:16531994 PMID:16798881

    Open questions at the time
    • Did not define how GINS/CDC45 mechanistically activate MCM
    • Strand handling within the helicase unknown
  4. 2010 High

    Showed mechanistically that CDC45 and GINS association activates the MCM ATPase and helicase, quantifying the activation and demonstrating cooperative complex formation.

    Evidence Reconstitution from recombinant Drosophila proteins with ATPase, helicase, and pulldown assays

    PMID:20122406

    Open questions at the time
    • Structural basis of activation not yet visualized
    • Did not address leading vs lagging strand routing
  5. 2011 High

    Provided the structural explanation for CMG activation, showing GINS and CDC45 bridge the Mcm2/Mcm5 gate to close the helicase ring.

    Evidence Single-particle EM of MCM2-7 and CMG with functional interpretation

    PMID:21378962

    Open questions at the time
    • Resolution insufficient for atomic detail of CDC45
    • DNA path within channel not directly resolved
  6. 2013 Medium

    Defined CDC45 as an ssDNA-binding RecJ/DHH-fold protein that engages 3'-protruding and forked structures, linking its architecture to a strand-handling role.

    Evidence Recombinant human/yeast CDC45 with EMSA, AFM, SPR, SAXS binding assays and in vivo mutant phenotypes

    PMID:22147708 PMID:23382391 PMID:24293646

    Open questions at the time
    • Most DHH catalytic residues not conserved; no nuclease activity demonstrated
    • Single-lab biochemistry
  7. 2015 High

    Established that CDC45 guards the leading strand within the CMG, defining its role in strand exclusion during unwinding.

    Evidence DNA-protein crosslinking, site-directed mutagenesis, and helicase assays on Drosophila CMG

    PMID:25561522

    Open questions at the time
    • Physiological consequence of leading-strand guarding in vivo not fully established
  8. 2016 High

    Resolved the atomic structure of human CDC45 and connected disease-relevant residues to CMG assembly mechanism.

    Evidence X-ray crystallography at 2.1 A with structure-guided mutagenesis and EM integration

    PMID:27189187

    Open questions at the time
    • Conformational changes upon CMG incorporation not captured
    • Function of the helical insertion only inferred
  9. 2021 High

    Defined how DDK phosphorylation of MCM tails templates a two-stage CMG assembly via Cdc45-tail-GINS intermediates, answering how CDC45 recruitment is regulated kinetically.

    Evidence Single-molecule fluorescence reconstitution with DDK phosphorylation (building on Sld3/GINS competition and ordered-loading studies)

    PMID:16990792 PMID:21362622 PMID:25659432 PMID:33616038

    Open questions at the time
    • The inefficient second step's molecular determinants remain undefined
    • In vivo correspondence of CtG intermediates not directly shown
  10. 2017 Medium

    Showed CDC45 catalytically loads RPA onto nascent ssDNA, extending its role beyond unwinding to coordinating ssDNA protection at the fork.

    Evidence Pull-down, SPR, and real-time RPA-ssDNA binding assays with human recombinant proteins

    PMID:10518787 PMID:17573775 PMID:28100698

    Open questions at the time
    • Catalytic mechanism of RPA handoff not structurally resolved
    • Single-lab kinetic study
  11. 2018 Medium

    Identified CDC45 as a checkpoint scaffold recruiting Rad53/checkpoint kinase and linked a CDC45 disordered loop to fork stabilization and a disease mutation.

    Evidence Co-IP, in vitro phosphorylation, genetic epistasis, and Meier-Gorlin mutation analysis (with Chk1/Chk2 displacement studies)

    PMID:16912045 PMID:22853956 PMID:30595439

    Open questions at the time
    • How checkpoint engagement at CDC45 is timed relative to elongation unclear
    • Single-lab Co-IP for several interactions
  12. 2016 Medium

    Linked CDC45 dosage to genome stability and established disease causation, showing CDC45 is rate-limiting for origin firing and causative for Meier-Gorlin syndrome.

    Evidence DNA fiber and cellular assays under CDC45 overexpression, plus whole-exome sequencing and patient-cell functional validation, with KO mouse lethality

    PMID:26919204 PMID:27374770 PMID:53

    Open questions at the time
    • Tissue-specific basis of MGS/craniosynostosis phenotype unexplained
    • Quantitative link between CDC45 level and origin number incompletely defined

Open questions

Synthesis pass · forward-looking unresolved questions
  • How CDC45 abundance, post-translational regulation (PP2A, ubiquitin/APC-C, DNAJA1 chaperone, Myc-driven recruitment) and checkpoint signaling are integrated to set origin firing rates and protect forks across differentiated tissues remains unresolved.
  • No unified model of CDC45 dosage control in vivo
  • Direct ubiquitylation and APC/C targeting not biochemically demonstrated
  • Tissue specificity of regulation unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 4 GO:0140097 catalytic activity, acting on DNA 4 GO:0003677 DNA binding 3 GO:0098772 molecular function regulator activity 2
Localization
GO:0000228 nuclear chromosome 3 GO:0005634 nucleus 3 GO:0005654 nucleoplasm 2
Pathway
R-HSA-69306 DNA Replication 4 R-HSA-1640170 Cell Cycle 3 R-HSA-8953897 Cellular responses to stimuli 3
Partners
DONSONGINS (PSF2)MCM10MCM7POLA1RPASLD3TOPBP1
Complex memberships
CMG helicase (Cdc45/Mcm2-7/GINS)Cdc45-MCM2-7-Sld3 (CMS) intermediateReplisome progression complex (RPC)

Evidence

Reading pass · 54 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2006 CDC45, MCM2-7, and GINS form a stable replisome progression complex (RPC) at eukaryotic replication forks. GINS is essential for maintaining the association of CDC45 with MCM within RPCs after initiation, and RPCs also contain Mrc1, Tof1-Csm3, FACT, Ctf4, Mcm10, and topoisomerase I. Immunoaffinity purification, mass spectrometry, chromatin immunoprecipitation (budding yeast) Nature cell biology High 16531994
2006 CDC45, MCM2-7, and GINS form the CMG (Cdc45/Mcm2-7/GINS) complex, which is the eukaryotic replicative DNA helicase. The purified complex from Drosophila embryo extracts has ATP-dependent DNA helicase activity. RNAi knockdown of GINS or CDC45 blocks S-phase transition. Immunoaffinity purification, in vitro helicase assay, RNAi knockdown (Drosophila) Proceedings of the National Academy of Sciences of the United States of America High 16798881
2010 Association of CDC45 and GINS with MCM2-7 activates the helicase: ATP hydrolysis rates are elevated ~100-fold, helicase activity is robust on circular templates, and DNA affinity is improved. GINS binds specifically to MCM4. All pairwise associations among GINS, MCMs, and CDC45 are detectable but tight association requires the full CMG. Reconstitution with recombinant Drosophila proteins, ATPase assay, in vitro helicase assay, pulldown Molecular cell High 20122406
2011 Cryo-EM structures of MCM2-7 and the CMG complex reveal that GINS and CDC45 bridge the Mcm2/Mcm5 gap in the helicase ring, forming a topologically closed assembly. Nucleotide binding further seals the ring, partitioning the central channel into two pores. This explains how GINS and CDC45 activate Mcm2-7 helicase. Single-particle electron microscopy, structural analysis Nature structural & molecular biology High 21378962
2016 Crystal structure of human CDC45 at 2.1 Å confirms evolutionary relationship to bacterial RecJ nuclease (DHH family). Key features include: long-range N-C terminal DHH domain interaction blocking the DNA-binding groove, and a helical insertion poised for replisome interactions. Mutational analysis validated the mechanism of CDC45 association with the MCM ring and GINS co-activator critical for CMG assembly. X-ray crystallography (2.1 Å), structure-guided mutagenesis, EM data integration Nature communications High 27189187
2000 CDC45 is required for origin unwinding during replication initiation in Xenopus. CDC45 binds chromatin upstream of RPA and DNA polymerase alpha. When CDC45 is present but DNA pol alpha is inhibited, helicase activity becomes uncoupled, demonstrating that CDC45 drives the unwinding step. Xenopus egg extract replication assay, immunodepletion, supercoiling assay, chromatin fractionation Molecular cell High 10882098
2006 At paused replication forks in Xenopus, MCM2-7, CDC45, and GINS are enriched at the unwinding site even when polymerase is inhibited by aphidicolin, establishing these three as core components of the 'unwindosome' that separates DNA strands at the replication fork. Biotin-streptavidin fork-pausing assay, chromatin immunoprecipitation, Xenopus egg extracts Molecular cell High 16483939
2004 MCM7 and CDC45 are required throughout replication elongation (not just initiation) in vertebrates. Antibody neutralization of CDC45 or MCM7 after significant DNA synthesis had already occurred blocked further synthesis and abolished helicase-dependent chromosome unwinding (uncoupled by aphidicolin), establishing CDC45 as a helicase co-factor essential for elongation. Xenopus egg extract, antibody neutralization, aphidicolin uncoupling assay The EMBO journal High 15329670
1998 Xenopus CDC45 loads DNA polymerase alpha onto chromatin at replication initiation. CDC45 physically interacts with DNA polymerase alpha in egg extracts, associates with chromatin only after nuclear formation in an S-phase CDK-dependent manner, and co-localizes with pol alpha in S-phase nuclei. Xenopus egg extract, co-immunoprecipitation, chromatin fractionation, immunofluorescence The EMBO journal High 9755170
2000 CDC45 is essential for the sequential chromatin loading of RPA, DNA pol alpha, and PCNA at replication initiation in Xenopus. CDC45 forms a stable complex with either MCM or DNA pol alpha on chromatin. DNA pol epsilon loading requires CDC45 but not pol alpha, suggesting a dual role in DNA unwinding and polymerase recruitment. Xenopus egg extract, immunodepletion, chromatin fractionation, co-immunoprecipitation Genes to cells : devoted to molecular & cellular mechanisms High 10886370
2001 CDC45 forms a complex with Sld3 throughout the cell cycle in S. cerevisiae. Their origin associations are mutually dependent. In sld3 mutants, the Sld3-CDC45 interaction and the CDC45-MCM2 interaction are both reduced. RPA does not associate with origins in the absence of Sld3, showing that the Sld3-CDC45 complex is prerequisite for origin unwinding. Co-immunoprecipitation, chromatin immunoprecipitation, two-hybrid, genetic analysis (budding yeast) The EMBO journal High 11296242
2012 The human CMG complex (CDC45/MCM2-7/GINS) purified from baculovirus-infected Sf9 cells has DNA helicase activity that: requires forked DNA structures for maximal activity; translocates 3' to 5' on the leading strand template; unwinds up to 500 bp; and, together with DNA pol epsilon, supports leading-strand synthesis >10 kb. Baculovirus expression, in vitro helicase assay, ATPase assay, rolling circle DNA synthesis assay (human CMG) Proceedings of the National Academy of Sciences of the United States of America High 22474384
2015 CDC45 guards the leading strand within the CMG: cross-linking studies show the leading strand contacts CDC45 when the MCM2/5 gate is open, but the lagging strand does not pass through the side channel. Mutations in the RecJ-like fold of CDC45 that ablate this leading-strand interaction diminish helicase activity. DNA-protein crosslinking, site-directed mutagenesis, in vitro helicase assay (Drosophila CMG) Proceedings of the National Academy of Sciences of the United States of America High 25561522
2011 CDC45 binds single-stranded DNA (ssDNA) with a structure similar to RecJ, demonstrating evolutionary relationship to DHH phosphoesterase family. Biochemical and SAXS data confirm only a subset of the Mn2+-coordinating residues are conserved, but the protein retains ssDNA (not dsDNA) binding activity. Recombinant human CDC45, SAXS, ssDNA binding assays, bioinformatics The Journal of biological chemistry Medium 22147708
2013 Human CDC45 binds long ssDNA (≥40 nt) and preferentially binds 3'-protruding strands, Y-shaped DNA, bubbles, and D-loops with higher affinity than short oligonucleotides. CDC45 slides on DNA with 3'-5' polarity, suggesting it acts as a molecular wedge to initiate strand displacement. Recombinant human CDC45, EMSA, AFM, SPR, SRCD, SAXS Nucleic acids research Medium 24293646
2013 CDC45 from budding yeast binds tightly to long (≥40 nt) ssDNA; 60-mer ssDNA disrupts the CDC45-MCM2-7 interaction. A CDC45 mutant unable to bind ssDNA causes helicase uncoupling from the polymerase under replication stress (hydroxyurea), with excess RPA accumulating near origins, demonstrating that CDC45-ssDNA interaction is required to stall the helicase during replication stress. Purified protein binding assays, site-directed mutagenesis, yeast genetics, chromatin immunoprecipitation The Journal of biological chemistry Medium 23382391
2017 Human CDC45 actively loads RPA onto nascent ssDNA in a catalytic manner. CDC45 forms a complex with RPA and stabilizes the 8-10 nt RPA binding mode; interaction requires the RPA70A subdomain. RPA dissociates when it covers a 30-mer. CDC45 facilitates ordered RPA deposition on ssDNA at the replication fork. Pull-down assay, surface plasmon resonance, real-time RPA-ssDNA binding assay (human recombinant proteins) Nucleic acids research Medium 28100698
2012 Checkpoint kinase Chk2 (but not Chk1) directly inhibits CMG helicase activity in vitro by phosphorylating MCM3, MCM4, and GINS subunit Psf2. Phosphatase treatment of CMG stimulates helicase activity. Ionizing radiation in Drosophila embryos causes hyperphosphorylation of Psf2 within the active helicase complex in vivo. In vitro kinase assay, helicase assay with recombinant Drosophila CMG, mass spectrometry, Drosophila embryo irradiation Proceedings of the National Academy of Sciences of the United States of America High 22853956
1997 CDC45 functions in late G1 phase after START and prior to DNA synthesis to trigger initiation at replication origins. CDC45 and CDC7/Dbf4 kinase are mutually dependent for function; cells defective in CDC45 cannot activate prereplicative complexes. Yeast genetics, cell cycle synchronization, epistasis analysis Proceedings of the National Academy of Sciences of the United States of America Medium 9356482
1997 CDC45 is essential for DNA replication initiation in S. cerevisiae. It genetically interacts with MCM genes (CDC46, CDC47, CDC54) and is synthetically lethal with orc2-1, mcm2-1, and mcm3-1. Origins fire less frequently in cdc45-1 cells, establishing CDC45 as functioning with ORC and MCM proteins in replication initiation. Yeast genetics, complementation, 2D gel origin firing analysis, synthetic lethality Molecular and cellular biology High 9001208
2006 Cdc7-Dbf4 kinase (DDK)-dependent phosphorylation of MCM4 N-terminal residues stimulates CDC45 association with chromatin. Deletion of MCM4 N-terminal 150 aa causes growth inhibition, and combined alanine substitution/deletion of N-terminal segments of MCM2, MCM4, and MCM6 leads to non-viable phenotype, indicating redundant but essential roles for these DDK-target sites in CDC45 loading. Chromatin fractionation, phospho-specific antibodies, SDS-PAGE mobility shift, Cdc7 conditional KO mouse ES cells, siRNA, mutagenesis (mammalian cells) The Journal of biological chemistry Medium 17046832
2002 Xenopus MCM10 binds chromatin after MCM2-7 but upstream of CDC45. In the absence of MCM10, CDC45 binding, RPA binding, and origin unwinding (supercoiling) are all blocked, placing MCM10 as an essential intermediate between pre-RC assembly and CDC45 loading. Xenopus egg extract, immunodepletion, chromatin fractionation, supercoiling assay Molecular cell High 11864598
2002 Protein phosphatase 2A (PP2A) is required for CDC45 loading onto the pre-RC in Xenopus. PP2A depletion or okadaic acid treatment abolishes CDC45 loading, origin unwinding, and downstream RPA and pol alpha loading. PP2A acts on a soluble factor (not CDC45 itself or pre-RC components) to enable CDC45 loading. Xenopus egg extract, PP2A immunodepletion, okadaic acid inhibition, chromatin fractionation The Journal of biological chemistry Medium 12185086
2002 Xmus101 (TOPBP1 ortholog) is required for loading CDC45 onto origins in Xenopus. Xmus101 chromatin association depends on ORC but is independent of MCM2-7 and S-CDK, defining a parallel ORC-dependent pathway for CDC45 loading distinct from the MCM2-7 pathway. Xenopus egg extract, immunodepletion, chromatin fractionation, epistasis The Journal of cell biology Medium 12438414
2011 Origin association timing of CDC45 (together with Sld3 and Sld7) is the key determinant of origin firing time in budding yeast. CDC45 associates with early-firing origins in G1 in a DDK-dependent manner; increased dosage of Sld3/Sld7/CDC45 allows late origins to fire earlier, as does increased DDK dosage. Chromatin immunoprecipitation, dosage analysis, genetic overexpression (budding yeast) Current biology : CB Medium 22169533
2006 In fission yeast, Sld3 is loaded at origins upstream of GINS, Cut5, and CDC45. DDK but not CDK is required for Sld3 loading, while CDC45 loading requires both kinases. GINS integrity is required for CDC45 loading but not Sld3 loading, establishing the ordered assembly: Sld3 → GINS/Cut5 → CDC45. Chromatin immunoprecipitation, temperature-sensitive mutant analysis, pull-down (fission yeast) The EMBO journal Medium 16990792
2009 Assembly of the human CMG complex (CDC45-MCM2-7-GINS) in HeLa cells requires CDK activity, CDC7 kinase, and the additional proteins RecQL4, Ctf4/And-1, and Mcm10. CMG interactions are only observed after G1/S transition. Depletion of TopBP1 did not significantly affect CMG complex formation in human cells. Bimolecular fluorescence complementation (BiFC) in HeLa cells, siRNA depletion, CDK inhibitor treatment Proceedings of the National Academy of Sciences of the United States of America Medium 19805216
2006 Chk1-mediated S-phase checkpoint targets CDC45 via a Cdc25A/CDK2-independent mechanism. BPDE-induced DNA damage causes Chk1-dependent reduction of chromatin-associated CDC45 (not soluble CDC45) and disrupts the CDC45-MCM7 interaction at the beta-globin replication origin, without affecting MCM7, MCM10, or PCNA chromatin binding. Chromatin fractionation, co-immunoprecipitation, chromatin immunoprecipitation, Chk1 inhibitor UCN-01 (human cells) The Journal of biological chemistry Medium 16912045
2018 CDC45 targets checkpoint kinase Rad53 to replication complexes via FHA-domain interaction with phosphorylated motifs in an intrinsically disordered loop of Cdc45. This interaction is necessary for Rad53-mediated inhibition of origin firing through Sld3, and also for stabilizing stalled forks. A CDC45 mutation found in Meier-Gorlin syndrome disrupts the Rad53 interaction. Co-immunoprecipitation, in vitro phosphorylation, genetic epistasis, budding yeast Molecular cell Medium 30595439
2011 GINS and Sld3 compete with each other for binding to both MCM2-7 and CDC45. Purified proteins form either a Cdc45-MCM2-7-Sld3 (CMS) or Cdc45-MCM2-7-GINS (CMG) complex with 1:1:1 stoichiometry. The data suggest GINS displaces Sld3 at the origin to activate the replication fork helicase. Purified recombinant protein binding assays, size exclusion chromatography, competition assays (budding yeast) The Journal of biological chemistry Medium 21362622
2021 DDK regulates CMG formation via a two-stage mechanism: DDK phosphorylation of MCM2-7 N-terminal tails recruits Cdc45 and GINS to form Cdc45-tail-GINS (CtG) intermediates. Higher DDK phosphorylation increases CtG multiplicity per MCM2-7, and higher CtG numbers increase the frequency of CMG formation in a second, inefficient step. Single-molecule fluorescence microscopy, in vitro reconstitution, DDK phosphorylation assay eLife High 33616038
2013 Human CDC45 interacts with Claspin, RPA, and DNA polymerase delta maximally during S phase. UV-induced DNA damage reduces CDC45-Claspin complex formation without affecting CDC45-RPA interaction, and this dissociation occurs upstream of ATR activation in the S-phase checkpoint. Co-immunoprecipitation, synchronized HeLa cells, UV treatment, kinase inhibitors The FEBS journal Low 23910567
2007 Human CDC45 co-localizes with active replication sites during S phase and interacts with DNA polymerase delta, DNA polymerase epsilon, GINS subunit Psf2, and MCM5/7 subunits, suggesting CDC45 bridges replicative polymerases with the MCM helicase in the elongation complex. Co-immunoprecipitation, immunofluorescence co-localization (human cells) Genes to cells : devoted to molecular & cellular mechanisms Low 17573775
1999 Human CDC45 directly binds hMCM7 and the p70 subunit of DNA polymerase alpha in vitro, supporting a role as molecular tether for loading pol alpha onto the replication complex via MCM7. In vitro binding assay (direct interaction), pull-down (human proteins) European journal of biochemistry Medium 10518787
1998 Human CDC45 (CDC45L) co-immunoprecipitates with human ORC2 from cell extracts; the protein associates with the nuclear fraction in G1 but this association becomes labile as S phase progresses, consistent with a role in replication initiation. Co-immunoprecipitation, subcellular fractionation (human cells) The Journal of biological chemistry Low 9660782
2005 Targeting CDC45 to specific chromosomal sites in mammalian cells induces large-scale chromatin decondensation correlated with histone H1 phosphorylation. CDC45 recruits CDK2 to these sites; CDK2 activity is required for decondensation. CDC45, CDK2, cyclin A, and phospho-H1 physically interact and associate with chromatin during S phase. Lac-repressor chromatin targeting, immunofluorescence, CDK2 inhibitors, co-immunoprecipitation (mammalian cells) The Journal of cell biology Medium 15753125
2008 Human TopBP1 directly interacts with CDC45 in vitro and in vivo, with this interaction occurring exclusively at the G1/S boundary. The first and second BRCT domains of TopBP1 mediate binding to CDC45, and overexpression of the sixth BRCT domain reduces CDC45 chromatin loading. GST pull-down, co-immunoprecipitation, deletion mutant analysis, yeast/mammalian one-hybrid, chromatin fractionation The Biochemical journal Medium 17887956
2010 DUE-B (DNA unwinding element binding protein) interacts with CDC45 and TopBP1 in cell extracts and baculovirus-expressed proteins. DUE-B and CDC45 co-localize at active replication origins. DUE-B immunodepletion in Xenopus egg extracts blocks replication and CDC45 (and a fraction of TopBP1) loading onto chromatin. Co-immunoprecipitation, baculovirus co-expression, chromatin immunoprecipitation, Xenopus immunodepletion Molecular and cellular biology Medium 20065034
2013 Human Ctf4 (hCtf4) forms a complex with the CMG helicase in vitro (purified proteins), in Sf9 cells, and from HeLa chromatin. hCtf4 is a homodimer that acts as a platform linking pol alpha to CMG. The hCtf4-CMG complex retains helicase activity with greater salt resistance than CMG alone. Stability of hCtf4-CMG depends on interactions with multiple CMG components. In vitro binding with purified proteins, co-infection Sf9 cells, HeLa chromatin immunoprecipitation, helicase assay Proceedings of the National Academy of Sciences of the United States of America High 24255107
2001 Fission yeast Sna41 (CDC45 ortholog) facilitates loading of DNA pol alpha onto MCM proteins in vivo. Sna41 interacts with pol alpha throughout the cell cycle and with Mcm6 in chromatin fractions at G1-S. In a sna41 initiation-defective mutant, pol alpha does not interact with MCM6, establishing CDC45 as essential for pol alpha-MCM association. In vivo tagged protein co-immunoprecipitation, chromatin fractionation, fission yeast genetics The Journal of biological chemistry Medium 11344166
2003 Following replication initiation in Xenopus, CDC45 and all six MCM subunits form a tight complex on chromatin in a CDK-dependent manner. This MCM-CDC45 chromatin complex has DNA helicase activity, which requires both CDK activity and CDC45, providing direct evidence for CDC45 as a helicase co-factor in vivo. Xenopus egg extract, denaturing immunoprecipitation, chromatin immunoprecipitation with helicase assay Genes to cells : devoted to molecular & cellular mechanisms Medium 12581157
2012 Mcm10 plays a role in CMG helicase function independent of CMG assembly. In budding yeast, auxin-induced degradation of Mcm10 allows stable CMG assembly at origins, but subsequent CMG translocation, RPA loading, and intra-S checkpoint activation are severely diminished. Mcm10 chromatin association depends on S-CDK and CDC45. Auxin-inducible degron, chromatin immunoprecipitation, budding yeast Current biology : CB Medium 22032285
2007 CDC45 protein is ubiquitylated and degraded via the proteasome pathway during terminal differentiation of human cells. Proteasome inhibitors decelerate CDC45 loss during differentiation. Multiple putative destruction boxes and a KEN-box suggest CDC45 is an APC/C substrate. CDC45 is not cleaved during apoptosis. Proteasome inhibitor treatment, immunoblotting (human cells) Biochemical and biophysical research communications Low 17767920
2013 Overexpression of CDC45 in Xenopus recapitulates c-Myc-induced replication phenotypes: increased density of early-replicating origins, elevated replication fork stalling/collapse, and DNA damage. CDC45 and GINS function downstream of Myc in regulating replication initiation. Xenopus egg extract, DNA fiber analysis, immunofluorescence, epistasis via overexpression Cell reports Medium 23643534
2023 DONSON is required for CDC45 and GINS association with MCM2-7 (CMG assembly) during replication initiation in Xenopus egg extracts. DONSON interacts with the initiation factor TopBP1 in a CDK-dependent manner. DONSON also associates with the replisome during elongation. Xenopus egg extracts, immunodepletion, chromatin fractionation, co-immunoprecipitation Nucleic acids research Medium 37638758
2016 In human cells, CDC45 overexpression fires at least twice as many origins but causes ~2-fold reduced fork elongation rate, pronounced fork asymmetry, S-phase arrest, accumulation of long ssDNA stretches (replication catastrophe), and ATM/Chk2-mediated H2AX phosphorylation, consistent with CDC45 being rate-limiting for origin firing. DNA fiber assay, flow cytometry, immunofluorescence, overexpression in human cell lines Cell cycle (Georgetown, Tex.) Medium 26919204
2019 Myc induces chromatin decondensation at targeted sites and directly promotes CDC45/GINS recruitment to resident MCMs, activating CMG helicases. Myc-Box II (MBII) and its interactors GCN5, Tip60, and TRRAP are required for chromatin unfolding and CDC45 recruitment. Myc and CDC45 physically interact. Co-immunoprecipitation, chromatin immunoprecipitation, lac-repressor chromatin targeting, siRNA depletion (mammalian cells) Communications biology Medium 30911685
2019 DNAJA1 (Hsp40 family) stabilizes CDC45 protein and promotes cell cycle progression. KNK437 reduces DNAJA1 levels, leading to reduced CDC45 stability. E2F1 transcriptionally activates DNAJA1, which then stabilizes CDC45 to promote the cell cycle. siRNA knockdown, Western blotting, co-immunoprecipitation (human colorectal cancer cells) Oncogene Low 31477839
2019 CDC45 contains an initiation-specific function: temperature-sensitive CDC45 mutants (in the RecJ-like domain and IDR) are defective for CMG formation and replication initiation but not elongation. The IDR of CDC45 is required for its function when carrying lethal point mutations but CDC45 lacking the IDR entirely retains full function, indicating the IDR context matters for initiation. Site-directed mutagenesis, temperature-sensitive yeast genetics, CMG formation assay, in vivo replication assay (budding yeast) PloS one Medium 30913274
2015 Dpb11 (human TopBP1 ortholog) binds Mcm2-7 and competes with GINS for Mcm2-7 binding. Dpb11 can recruit CDC45 to Mcm2-7. ssDNA inhibits Dpb11-Mcm2-7 interaction, allowing GINS to displace Dpb11 and bind Mcm2-7 for CMG assembly. Purified protein binding assays, competition experiments, yeast genetics, ChIP (budding yeast) The Journal of biological chemistry Medium 25659432
2013 Human CDC45 directly interacts with all MCM2-7 subunits and with PSF2, PSF3, and SLD5 (GINS subunits), as well as RPA2, AND-1, and TopBP1 by immunoprecipitation. A considerable portion of CDC45 in nuclei is associated with nuclear scaffold structures (nuclease-resistant fraction) rather than at replication forks. Immunoprecipitation, chromatin fractionation, nuclease treatment, synchronized HeLa cells Journal of biochemistry Low 23364835
2019 PP2A exists in complex with CDC45 during DNA replication, and increased PP2A activity causes dissociation of CDC45 and polymerase alpha from the replisome, interrupting ongoing DNA replication and causing replication fork collapse. Co-immunoprecipitation, PP2A activator (small molecule), PP2A genetic loss-of-function, DNA replication assays (human cells) The Journal of biological chemistry Medium 31562245
2016 Biallelic partial loss-of-function mutations in human CDC45 cause Meier-Gorlin syndrome and craniosynostosis. Mutations reduce full-length CDC45 transcript and protein levels in patient cells, consistent with reduced DNA replication rate and cell proliferation. CDC45 is thus functionally distinct from pre-RC MGS genes, implicating the pre-IC in MGS etiology. Whole-exome sequencing, RT-PCR splicing analysis, patient cell lines (protein quantification) American journal of human genetics Medium 27374770
2001 CDC45 null mouse embryos fail to develop past implantation; inner cell mass shows impaired proliferation, establishing CDC45 as essential for mammalian post-implantation development and cell proliferation in vivo. Heterozygous mice develop normally, suggesting hemizygosity of CDC45 alone is insufficient to cause cardiac/craniofacial defects in DiGeorge syndrome. Gene targeting (knockout mouse), embryo culture, immunostaining Molecular and cellular biology Medium 11416137

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2006 GINS maintains association of Cdc45 with MCM in replisome progression complexes at eukaryotic DNA replication forks. Nature cell biology 618 16531994
2006 Isolation of the Cdc45/Mcm2-7/GINS (CMG) complex, a candidate for the eukaryotic DNA replication fork helicase. Proceedings of the National Academy of Sciences of the United States of America 564 16798881
2010 Activation of the MCM2-7 helicase by association with Cdc45 and GINS proteins. Molecular cell 456 20122406
2000 Initiation of eukaryotic DNA replication: origin unwinding and sequential chromatin association of Cdc45, RPA, and DNA polymerase alpha. Molecular cell 335 10882098
2006 Localization of MCM2-7, Cdc45, and GINS to the site of DNA unwinding during eukaryotic DNA replication. Molecular cell 306 16483939
2011 The structural basis for MCM2-7 helicase activation by GINS and Cdc45. Nature structural & molecular biology 270 21378962
2004 A requirement for MCM7 and Cdc45 in chromosome unwinding during eukaryotic DNA replication. The EMBO journal 207 15329670
2011 Origin association of Sld3, Sld7, and Cdc45 proteins is a key step for determination of origin-firing timing. Current biology : CB 202 22169533
1998 Xenopus Cdc45-dependent loading of DNA polymerase alpha onto chromatin under the control of S-phase Cdk. The EMBO journal 195 9755170
2001 Sld3, which interacts with Cdc45 (Sld4), functions for chromosomal DNA replication in Saccharomyces cerevisiae. The EMBO journal 186 11296242
2009 Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins. Proceedings of the National Academy of Sciences of the United States of America 162 19805216
2002 Xenopus Mcm10 binds to origins of DNA replication after Mcm2-7 and stimulates origin binding of Cdc45. Molecular cell 158 11864598
2006 Phosphorylation of MCM4 by Cdc7 kinase facilitates its interaction with Cdc45 on the chromatin. The Journal of biological chemistry 154 17046832
1997 CDC45, a novel yeast gene that functions with the origin recognition complex and Mcm proteins in initiation of DNA replication. Molecular and cellular biology 142 9001208
2013 Cdc45 is a critical effector of myc-dependent DNA replication stress. Cell reports 113 23643534
2012 Properties of the human Cdc45/Mcm2-7/GINS helicase complex and its action with DNA polymerase epsilon in rolling circle DNA synthesis. Proceedings of the National Academy of Sciences of the United States of America 113 22474384
2000 Central role for cdc45 in establishing an initiation complex of DNA replication in Xenopus egg extracts. Genes to cells : devoted to molecular & cellular mechanisms 113 10886370
2002 The Xenopus Xmus101 protein is required for the recruitment of Cdc45 to origins of DNA replication. The Journal of cell biology 111 12438414
2016 Mutations in CDC45, Encoding an Essential Component of the Pre-initiation Complex, Cause Meier-Gorlin Syndrome and Craniosynostosis. American journal of human genetics 96 27374770
2012 The CMG (CDC45/RecJ, MCM, GINS) complex is a conserved component of the DNA replication system in all archaea and eukaryotes. Biology direct 95 22329974
2005 Chromatin decondensation in S-phase involves recruitment of Cdk2 by Cdc45 and histone H1 phosphorylation. The Journal of cell biology 95 15753125
2012 Mcm10 plays a role in functioning of the eukaryotic replicative DNA helicase, Cdc45-Mcm-GINS. Current biology : CB 93 22285032
2003 Fission yeast Cdc23/Mcm10 functions after pre-replicative complex formation to promote Cdc45 chromatin binding. Molecular biology of the cell 89 12972571
2009 The human GINS complex associates with Cdc45 and MCM and is essential for DNA replication. Nucleic acids research 87 19223333
1997 CDC45 is required in conjunction with CDC7/DBF4 to trigger the initiation of DNA replication. Proceedings of the National Academy of Sciences of the United States of America 86 9356482
2015 Cdc45 (cell division cycle protein 45) guards the gate of the Eukaryote Replisome helicase stabilizing leading strand engagement. Proceedings of the National Academy of Sciences of the United States of America 79 25561522
2006 The Chk1-mediated S-phase checkpoint targets initiation factor Cdc45 via a Cdc25A/Cdk2-independent mechanism. The Journal of biological chemistry 79 16912045
2011 Cdc45 limits replicon usage from a low density of preRCs in mammalian cells. PloS one 76 21390258
1999 A role for the replication proteins PCNA, RF-C, polymerase epsilon and Cdc45 in transcriptional silencing in Saccharomyces cerevisiae. Genetics 75 10545450
2006 Ordered assembly of Sld3, GINS and Cdc45 is distinctly regulated by DDK and CDK for activation of replication origins. The EMBO journal 72 16990792
2016 Structure of human Cdc45 and implications for CMG helicase function. Nature communications 70 27189187
2004 Mcm10 and Cdc45 cooperate in origin activation in Saccharomyces cerevisiae. Journal of molecular biology 66 15201046
2011 Cdc45: the missing RecJ ortholog in eukaryotes? Bioinformatics (Oxford, England) 65 21653514
2003 CDK- and Cdc45-dependent priming of the MCM complex on chromatin during S-phase in Xenopus egg extracts: possible activation of MCM helicase by association with Cdc45. Genes to cells : devoted to molecular & cellular mechanisms 65 12581157
2007 Human Cdc45 is a proliferation-associated antigen. The FEBS journal 62 17608804
2016 Cdc45 is limiting for replication initiation in humans. Cell cycle (Georgetown, Tex.) 61 26919204
2007 Interactions of human Cdc45 with the Mcm2-7 complex, the GINS complex, and DNA polymerases delta and epsilon during S phase. Genes to cells : devoted to molecular & cellular mechanisms 57 17573775
2019 KNK437 restricts the growth and metastasis of colorectal cancer via targeting DNAJA1/CDC45 axis. Oncogene 56 31477839
2006 Cdc45-MCM-GINS, a new power player for DNA replication. Cell division 56 16930479
2017 Combined inhibition of Wee1 and Chk1 gives synergistic DNA damage in S-phase due to distinct regulation of CDK activity and CDC45 loading. Oncotarget 50 28030798
2011 Structural and functional insights into the DNA replication factor Cdc45 reveal an evolutionary relationship to the DHH family of phosphoesterases. The Journal of biological chemistry 50 22147708
2009 H3 k36 methylation helps determine the timing of cdc45 association with replication origins. PloS one 47 19521516
2019 Analysis of functional hub genes identifies CDC45 as an oncogene in non-small cell lung cancer - a short report. Cellular oncology (Dordrecht, Netherlands) 46 30887286
1998 The human homolog of Saccharomyces cerevisiae CDC45. The Journal of biological chemistry 46 9660782
2011 The Cdc45·Mcm2-7·GINS protein complex in trypanosomes regulates DNA replication and interacts with two Orc1-like proteins in the origin recognition complex. The Journal of biological chemistry 45 21799014
2010 The DNA unwinding element binding protein DUE-B interacts with Cdc45 in preinitiation complex formation. Molecular and cellular biology 45 20065034
1999 CDC45 and DPB11 are required for processive DNA replication and resistance to DNA topoisomerase I-mediated DNA damage. Proceedings of the National Academy of Sciences of the United States of America 45 10500195
1999 Human CDC45 protein binds to minichromosome maintenance 7 protein and the p70 subunit of DNA polymerase alpha. European journal of biochemistry 44 10518787
2013 Interaction between human Ctf4 and the Cdc45/Mcm2-7/GINS (CMG) replicative helicase. Proceedings of the National Academy of Sciences of the United States of America 43 24255107
2016 Archaeal orthologs of Cdc45 and GINS form a stable complex that stimulates the helicase activity of MCM. Proceedings of the National Academy of Sciences of the United States of America 41 27821767
2009 Regulation of Cdc45 in the cell cycle and after DNA damage. Biochemical Society transactions 37 19614620
2008 Apoptosis induced by replication inhibitors in Chk1-depleted cells is dependent upon the helicase cofactor Cdc45. Cell death and differentiation 36 18239674
2002 Protein phosphatase 2A regulates binding of Cdc45 to the prereplication complex. The Journal of biological chemistry 35 12185086
2012 Checkpoint kinase 2 (Chk2) inhibits the activity of the Cdc45/MCM2-7/GINS (CMG) replicative helicase complex. Proceedings of the National Academy of Sciences of the United States of America 34 22853956
2013 Cdc45 protein-single-stranded DNA interaction is important for stalling the helicase during replication stress. The Journal of biological chemistry 33 23382391
2013 DNA binding properties of human Cdc45 suggest a function as molecular wedge for DNA unwinding. Nucleic acids research 32 24293646
2001 Requirement of CDC45 for postimplantation mouse development. Molecular and cellular biology 32 11416137
2001 Essential role of Sna41/Cdc45 in loading of DNA polymerase alpha onto minichromosome maintenance proteins in fission yeast. The Journal of biological chemistry 31 11344166
2008 Characterization of the interaction between the human DNA topoisomerase IIbeta-binding protein 1 (TopBP1) and the cell division cycle 45 (Cdc45) protein. The Biochemical journal 30 17887956
2023 DONSON facilitates Cdc45 and GINS chromatin association and is essential for DNA replication initiation. Nucleic acids research 29 37638758
2021 DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7. eLife 29 33616038
2010 Hsk1 kinase and Cdc45 regulate replication stress-induced checkpoint responses in fission yeast. Cell cycle (Georgetown, Tex.) 29 21099360
2018 Helicase Subunit Cdc45 Targets the Checkpoint Kinase Rad53 to Both Replication Initiation and Elongation Complexes after Fork Stalling. Molecular cell 28 30595439
2017 The Cdc45/RecJ-like protein forms a complex with GINS and MCM, and is important for DNA replication in Thermococcus kodakarensis. Nucleic acids research 26 28977567
2017 Cdc45-induced loading of human RPA onto single-stranded DNA. Nucleic acids research 24 28100698
2011 GINS and Sld3 compete with one another for Mcm2-7 and Cdc45 binding. The Journal of biological chemistry 24 21362622
2019 Myc-driven chromatin accessibility regulates Cdc45 assembly into CMG helicases. Communications biology 23 30911685
2004 Schizosaccharomyces pombe replication protein Cdc45/Sna41 requires Hsk1/Cdc7 and Rad4/Cut5 for chromatin binding. Chromosoma 22 15338237
2019 Dynamic relocalization of replication origins by Fkh1 requires execution of DDK function and Cdc45 loading at origins. eLife 21 31084713
2023 IGF2BP2 promotes glycolysis and hepatocellular carcinoma stemness by stabilizing CDC45 mRNA via m6A modification. Cell cycle (Georgetown, Tex.) 19 37985379
2015 Dpb11 protein helps control assembly of the Cdc45·Mcm2-7·GINS replication fork helicase. The Journal of biological chemistry 19 25659432
2015 Human DNA helicase B interacts with the replication initiation protein Cdc45 and facilitates Cdc45 binding onto chromatin. Experimental cell research 19 25933514
2013 Protein interaction and cellular localization of human CDC45. Journal of biochemistry 19 23364835
2013 The physical interaction of Mcm10 with Cdc45 modulates their DNA-binding properties. The Biochemical journal 18 23750504
2019 Pathogenic variants in CDC45 on the remaining allele in patients with a chromosome 22q11.2 deletion result in a novel autosomal recessive condition. Genetics in medicine : official journal of the American College of Medical Genetics 17 31474763
2017 The crystal structure of Pyrococcus furiosus RecJ implicates it as an ancestor of eukaryotic Cdc45. Nucleic acids research 17 30053256
2016 TIMELESS Suppresses the Accumulation of Aberrant CDC45·MCM2-7·GINS Replicative Helicase Complexes on Human Chromatin. The Journal of biological chemistry 17 27587400
2022 Systematic pan‑cancer analysis identifies CDC45 as having an oncogenic role in human cancers. Oncology reports 15 36082823
2001 Sna41goa1, a novel mutation causing G1/S arrest in fission yeast, is defective in a CDC45 homolog and interacts genetically with polalpha. Molecular genetics and genomics : MGG 15 11523776
2008 Human CDT1 associates with CDC7 and recruits CDC45 to chromatin during S phase. The Journal of biological chemistry 14 19054765
2021 Knockdown of circular RNA hsa_circ_0062270 suppresses the progression of melanoma via downregulation of CDC45. Histology and histopathology 13 34931691
2013 Cell cycle-dependent formation of Cdc45-Claspin complexes in human cells is compromized by UV-mediated DNA damage. The FEBS journal 13 23910567
1998 Direct selection of conserved cDNAs from the DiGeorge critical region: isolation of a novel CDC45-like gene. Genome research 13 9724329
2022 High abundance of CDC45 inhibits cell proliferation through elevation of HSPA6. Cell proliferation 12 35642733
2021 A synonymous variant in a non-canonical exon of CDC45 disrupts splicing in two affected sibs with Meier-Gorlin syndrome with craniosynostosis. European journal of medical genetics 12 33639314
2020 RYBP inhibits esophageal squamous cell carcinoma proliferation through downregulating CDC6 and CDC45 in G1-S phase transition process. Life sciences 12 32209426
2019 Protein phosphatase 2A controls ongoing DNA replication by binding to and regulating cell division cycle 45 (CDC45). The Journal of biological chemistry 12 31562245
2012 Cell cycle-dependent mobility of Cdc45 determined in vivo by fluorescence correlation spectroscopy. PloS one 12 22536402
2020 DNA replication protein Cdc45 directly interacts with PCNA via its PIP box in Leishmania donovani and the Cdc45 PIP box is essential for cell survival. PLoS pathogens 11 32413071
1999 UFD1L and CDC45L: a role in DiGeorge syndrome and related phenotypes? Trends in genetics : TIG 11 10390621
2021 CDC45 modulates MCM7 expression and inhibits cell proliferation by suppressing the PI3K/AKT pathway in acute myeloid leukemia. American journal of translational research 10 34650692
2024 CDC45 promotes the stemness and metastasis in lung adenocarcinoma by affecting the cell cycle. Journal of translational medicine 8 38589907
2016 Structural insights into Cdc45 function: was there a nuclease at the heart of the ancestral replisome? Biophysical chemistry 8 27919598
2008 A kinetoplastid BRCA2 interacts with DNA replication protein CDC45. International journal for parasitology 8 18723021
2001 A common cis-acting sequence in the DiGeorge critical region regulates bi-directional transcription of UFD1L and CDC45L. Mechanisms of development 8 11578863
2007 Cdc45 degradation during differentiation and apoptosis. Biochemical and biophysical research communications 7 17767920
2001 Characterization of the bi-directional transcriptional control region between the human UFD1L and CDC45L genes. Biochemical and biophysical research communications 6 11341762
2024 Itaconate Ameliorates Experimental Autoimmune Uveitis by Modulating Teff/Treg Cell Imbalance Via the DNAJA1/CDC45 Axis. Investigative ophthalmology & visual science 5 39661355
2018 Cdc45/Mcm2-7/GINS complex down-regulation mediates S phase arrest in okadaic acid-induced cell damage. Toxicon : official journal of the International Society on Toxinology 5 30003918
2019 Initiation-specific alleles of the Cdc45 helicase-activating protein. PloS one 4 30913274

Missed literature

Know a paper Affinage missed for CDC45? Flag it for the maintainers and the community.

No submissions yet.