Affinage

MCM4

DNA replication licensing factor MCM4 · UniProt P33991

Length
863 aa
Mass
96.6 kDa
Annotated
2026-04-28
85 papers in source corpus 40 papers cited in narrative 40 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

MCM4 is an essential subunit of the MCM2-7 replicative helicase complex that functions at the core of eukaryotic DNA replication initiation and elongation. Within the MCM4/6/7 sub-complex, MCM4 contributes to a ring-shaped heterohexamer with intrinsic 3′→5′ DNA helicase activity that unwinds duplex DNA by a steric exclusion mechanism, with its N-terminal residues (Arg10/11) required for helicase function and its zinc finger mediating inter-subunit contacts (PMID:9305914, PMID:13679365, PMID:12207017, PMID:30184107). The unstructured N-terminal serine/threonine-rich domain (NSD) of MCM4 integrates CDK, DDK (Cdc7/Dbf4), and checkpoint kinase (ATR/Mec1) signals: CDK phosphorylation inhibits helicase activity and reduces chromatin affinity to prevent re-replication, DDK phosphorylation relieves intrinsic NSD-mediated inhibition to license S-phase entry and promote Cdc45 loading, and checkpoint phosphorylation independently controls late-origin firing and fork progression under replication stress (PMID:8901561, PMID:11454864, PMID:20054399, PMID:24740181, PMID:17046832). MCM2-7 ring closure at the Mcm2/Mcm5 gate triggers Mcm4-specific ATP hydrolysis to drive Cdt1 release during pre-replicative complex assembly, while the MCM4 C-terminal domain restrains excessive unwinding at stalled replication forks (PMID:39747125, PMID:18753627). Hypomorphic MCM4 mutations in humans cause genomic instability and natural killer cell deficiency, and destabilizing mutations in mice promote chromosomal instability and cancer (PMID:22354167, PMID:17143284, PMID:23133403).

Mechanistic history

Synthesis pass · year-by-year structured walk · 15 steps
  1. 1992 Medium

    Establishing that MCM4 orthologs are required for DNA replication origin function answered the fundamental question of whether MCM proteins act in replication initiation rather than another cell-cycle process.

    Evidence S. pombe cdc21 genetic analysis with plasmid maintenance and FACS DNA content assays

    PMID:1454522

    Open questions at the time
    • No biochemical activity demonstrated
    • Precise step in replication not defined
  2. 1996 High

    Demonstration that MCM4 binds chromatin in an underphosphorylated state and is displaced by Cdc2/cyclin B phosphorylation established the first mechanistic link between CDK activity and MCM regulation, explaining how re-replication is prevented after S phase.

    Evidence Xenopus egg extracts, in vitro cdc2/cyclinB kinase assay, chromatin binding assays, and phosphorylation site mapping; corroborated by human cell immunofluorescence showing MCM4 marks unreplicated chromatin

    PMID:8605878 PMID:8838654 PMID:8901561

    Open questions at the time
    • Specific phosphorylation sites mediating chromatin release not mapped
    • Helicase activity not yet measured
  3. 1997 High

    Identification of intrinsic 3′→5′ DNA helicase and ATPase activities in the purified MCM4/6/7 complex answered the long-standing question of whether MCM proteins constitute the replicative helicase.

    Evidence Biochemical purification from HeLa cells with anti-MCM4 immunodepletion; helicase displacement and ATPase assays

    PMID:9305914

    Open questions at the time
    • Role of individual subunits in catalysis not resolved
    • Full MCM2-7 helicase activity not reconstituted
  4. 1999 High

    Mutagenesis of Walker A/B motifs separated the contributions of MCM4 (ssDNA binding) and MCM6 (ATP hydrolysis critical for helicase) within the heterotrimer, and CDK-driven nuclear exclusion of Mcm4 in budding yeast established a second re-replication block mechanism.

    Evidence Baculovirus-expressed mouse MCM4/6/7 with ATPase motif mutations; live-cell imaging of GFP-Mcm4 with cyclin alleles in S. cerevisiae

    PMID:10559985 PMID:10567526

    Open questions at the time
    • Whether nuclear exclusion operates in metazoans unclear
    • Structural basis for ssDNA binding by MCM4 unknown
  5. 2000 High

    EM visualization of the MCM4/6/7 ring structure and demonstration that CDK2/cyclin A phosphorylation of MCM4's N-terminus inhibits helicase activity established the physical architecture and a direct regulatory mechanism for the complex.

    Evidence Negative-stain EM of ring structures; in vitro Cdk2/cyclin A phosphorylation coupled to helicase assay; Xenopus extract phosphorylation state analysis

    PMID:10748114 PMID:10779356 PMID:10884341

    Open questions at the time
    • Phosphorylation sites not yet mapped at residue resolution
    • How MCM2 and MCM3/5 inhibit MCM4/6/7 mechanistically undefined
  6. 2001 High

    Mapping six specific Ser/Thr CDK sites in the MCM4 N-terminus and showing their mutation relieves helicase inhibition answered which residues mediate CDK-dependent licensing control.

    Evidence Site-directed mutagenesis; in vitro helicase assay; anti-phosphothreonine antibody; HeLa cell-cycle fractionation

    PMID:11454864

    Open questions at the time
    • Whether all six sites are individually required or act combinatorially unclear
    • In vivo functional consequence of site mutations not tested in metazoans
  7. 2003 High

    Definition of the steric exclusion ('pump in ring') unwinding mechanism and discovery that MCM4/6/7 is activated by thymine-rich ssDNA sequences at bubble/fork structures linked helicase biochemistry to origin-specific activation.

    Evidence In vitro helicase assays with systematic bubble, Y-fork, and Holliday junction substrates; human lamin B2 origin sequences

    PMID:13679365 PMID:14609960

    Open questions at the time
    • Whether T-rich sequence preference operates in vivo at origins unknown
    • How full MCM2-7 complex modifies substrate specificity not tested
  8. 2003 High

    Linking ATR-CHK1 checkpoint signaling to sequential CDK2 phosphorylation of MCM4 under replication stress established MCM4 as a direct checkpoint effector that suppresses fork progression.

    Evidence Phospho-specific antibodies in HU/UV-treated HeLa cells; kinase inhibitor epistasis; in vitro helicase inhibition

    PMID:12714602

    Open questions at the time
    • Whether checkpoint phosphorylation of MCM4 is sufficient for fork arrest in vivo not shown
    • Direct ATR phosphorylation of MCM4 versus indirect CDK2-mediated effect not fully resolved
  9. 2006 High

    Identification of Cdc7/DDK phosphorylation of MCM4 N-terminal sites as a trigger for Cdc45 chromatin loading and cell-cycle-resolved mapping of seven individual phospho-sites connected specific kinase inputs to origin activation.

    Evidence Phospho-amino acid antibodies; Cdc7-null mouse ES cells; Cdc45 chromatin association; temperature-sensitive CDK1 mutant mouse cells

    PMID:16519687 PMID:17046832

    Open questions at the time
    • Structural mechanism by which DDK phosphorylation promotes Cdc45 recruitment unknown
    • Whether all seven sites are phosphorylated on the same molecule simultaneously unclear
  10. 2008 High

    The MCM4 C-terminal domain was identified as a brake that limits excessive unwinding at stalled forks, and S-CDK phosphorylation of MCM4 N-terminal sites was shown to cooperate with DDK for viability, establishing that MCM4 integrates both kinase inputs.

    Evidence S. pombe mcm4-c84 CTD truncation with ChIP and 2D gel analysis; S. cerevisiae mcm4-5A synthetic lethality with DDK gain-of-function alleles

    PMID:18321994 PMID:18753627

    Open questions at the time
    • Whether the CTD directly contacts DNA or acts through protein interactions unknown
    • Structural basis for CDK-DDK cooperation through MCM4 NSD not defined
  11. 2010 High

    Demonstration that DDK's sole essential function is relieving inhibition by the MCM4 NSD, and that all inter-subunit ATP sites in the MCM4/6/7 hexamer are required for helicase activity, defined the minimal catalytic unit and the key regulatory switch for S-phase entry.

    Evidence S. cerevisiae NSD deletion bypassing DDK essentiality; arginine finger mutagenesis of reconstituted Mcm4/6/7 with sedimentation and helicase assays

    PMID:20054399 PMID:20860810

    Open questions at the time
    • How the NSD physically inhibits the helicase not structurally resolved
    • Whether NSD bypass is conserved in metazoans unknown
  12. 2012 High

    Mouse and human genetic studies established that hypomorphic MCM4 mutations cause chromosomal instability, cancer predisposition, and NK cell deficiency, connecting MCM4 helicase function to genome maintenance and immune cell proliferation.

    Evidence Mouse Chaos3 (F345I) forward screen with mammary tumor phenotype; Sdl (D573H) dominant-negative mutation causing T-ALL; human splice-site mutation with NK cell deficiency rescued by WT MCM4 complementation

    PMID:17143284 PMID:22354167 PMID:23133403

    Open questions at the time
    • Why specific tissues (mammary, thymus, NK cells) are preferentially affected unclear
    • Whether human MCM4 mutations are common cancer drivers not established
  13. 2014 High

    Dissection of the MCM4 NSD into functionally independent proximal (checkpoint/late-origin) and distal (CDK/fork progression) segments resolved how a single domain integrates distinct kinase pathways to separately control origin timing and fork speed.

    Evidence Whole-genome replication profiling and single-molecule DNA fiber analysis with NSD segment-specific mutations in S. cerevisiae

    PMID:24740181

    Open questions at the time
    • Structural basis for sub-domain specificity not determined
    • Whether metazoan MCM4 NSD has analogous modular architecture unknown
  14. 2019 High

    A conserved Mcm4 motif was shown to be required for stable MCM2-7 double-hexamer formation, identifying this step as essential for extensive origin DNA unwinding even when helicase-activation factors are recruited.

    Evidence Single-molecule imaging of MCM loading and biochemical reconstitution with Mcm4 mutants in S. cerevisiae

    PMID:31385807

    Open questions at the time
    • Atomic contacts mediating double-hexamer interface through Mcm4 not fully resolved
    • Whether the motif functions identically in metazoan pre-RC assembly unknown
  15. 2025 High

    Cryo-EM structural analysis revealed that MCM2-7 ring closure at the Mcm2/Mcm5 gate specifically triggers Mcm4 ATP hydrolysis to drive Cdt1 release, identifying MCM4 as the catalytic trigger for pre-RC maturation.

    Evidence Cryo-EM of ORC-Cdc6-Cdt1-MCM2-7 intermediate; ATPase and ring-closure mutagenesis; biochemical reconstitution

    PMID:39747125

    Open questions at the time
    • How subsequent ATPase events by other MCM subunits coordinate with Mcm4 hydrolysis unclear
    • Whether ring closure sensing operates identically at all origins in vivo unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • The structural mechanism by which the MCM4 NSD physically inhibits helicase activity, and how DDK phosphorylation conformationally relieves this inhibition, remains unresolved at atomic resolution.
  • No high-resolution structure of the NSD in inhibitory or phosphorylated states
  • Whether NSD inhibition involves intra-molecular contacts with the AAA+ domain or inter-subunit contacts unknown
  • Metazoan conservation of the DDK bypass mechanism not tested

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016787 hydrolase activity 4 GO:0140657 ATP-dependent activity 4 GO:0003677 DNA binding 3
Localization
GO:0005694 chromosome 5 GO:0005634 nucleus 3
Pathway
R-HSA-1640170 Cell Cycle 6 R-HSA-69306 DNA Replication 6 R-HSA-73894 DNA Repair 4
Partners
CDC45CDC7CDT1MCM2MCM5MCM6MCM7
Complex memberships
MCM2-7 hexamerMCM4/6/7 sub-complexpre-replicative complex (pre-RC)

Evidence

Reading pass · 40 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1997 A DNA helicase activity is intrinsically associated with a 600-kDa complex of MCM4, MCM6, and MCM7 proteins. The complex also has ATPase activity, and immunodepletion with anti-MCM4 antibody severely reduces helicase activity. The helicase translocates along single-stranded DNA in the 3' to 5' direction and requires hydrolyzable ATP or dATP. Biochemical purification from HeLa cells, immunodepletion with anti-MCM4 antibody, ATPase and DNA helicase displacement assays The Journal of biological chemistry High 9305914
1999 The DNA helicase activity is intrinsic to the MCM4/6/7 complex. The ATP binding activity of MCM6 is critical for helicase activity, while MCM4 plays a role in single-stranded DNA binding activity of the complex. ATPase motif mutations in MCM4 and MCM6 enabled separation of helicase and ssDNA binding activities. Baculovirus expression of mouse Mcm2,4,6,7; Walker A/B motif mutagenesis; in vitro ATPase and helicase assays Molecular and cellular biology High 10567526
1996 Xenopus MCM4 is phosphorylated by cdc2/cyclin B kinase during mitosis at the same sites phosphorylated in vivo. This phosphorylation dramatically reduces MCM4 affinity for chromatin, providing a mechanism to inactivate the MCM complex from late S phase through mitosis and prevent illegitimate DNA re-replication. Cell-free Xenopus extract system, in vitro kinase assay with cdc2/cyclinB, chromatin binding assays, phosphorylation site mapping Proceedings of the National Academy of Sciences of the United States of America High 8901561
2000 Cyclin A/Cdk2 phosphorylates mainly the amino-terminal region of MCM4 within the MCM4/6/7 complex, and this phosphorylation is associated with inactivation of the DNA helicase activity of the complex. In vitro phosphorylation of purified MCM4/6/7 complex with cyclin A/Cdk2; helicase activity assay before and after phosphorylation The Journal of biological chemistry High 10748114
2001 Six Ser/Thr residues in the N-terminal region of MCM4 are required for CDK phosphorylation. Mutation of these sites largely relieves Cdk2/cyclin A-mediated inhibition of MCM4/6/7 helicase activity. One Thr site is phosphorylated specifically in mitosis, and MCM4/6/7 complexes purified from mitotic HeLa cells show reduced helicase activity correlating with phosphorylation at these sites. Site-directed mutagenesis of Mcm4 CDK sites; in vitro helicase assay; anti-phosphothreonine antibody; cell-cycle-phase fractionation of HeLa cells The Journal of biological chemistry High 11454864
2003 MCM4/6/7 is a ring-shaped heterohexamer that unwinds DNA by a 'pump in ring' steric exclusion mechanism, binding to only one strand (leading strand) of a duplex. The ring actively translocates along duplex DNA and can drive branch migration of Holliday junctions. Unwinding stops at a nick in either strand. In vitro helicase assays with various DNA substrates, strand-binding assays, Holliday junction branch migration assay The Journal of biological chemistry High 13679365
2003 Human MCM4 is extensively phosphorylated in response to DNA replication inhibitors (hydroxyurea) or UV irradiation via the sequential action of ATR-CHK1 and CDK2 kinases. CDK2 phosphorylation of MCM4 negatively regulates the DNA helicase activity of the MCM4-6-7 complex, suggesting that checkpoint-induced phosphorylation inhibits DNA fork progression. Phospho-specific antibodies against MCM4; kinase inhibition experiments; in vitro helicase activity of CDK2-phosphorylated MCM4/6/7 The Journal of biological chemistry High 12714602
2006 Cdc7 kinase phosphorylates MCM4 at specific N-terminal (S/T)(S/T)P residues during S phase on chromatin. This phosphorylation stimulates association of Cdc45 with chromatin and is required for origin activation. Deletion of the N-terminal 150 amino acids of MCM4 causes growth inhibition, and this is partially rescued by sequences containing Cdc7 target residues. Phospho-amino acid-specific antibodies; Cdc7-null mouse ES cells and siRNA knockdown; Cdc45 chromatin association assay; MCM4 deletion mutant complementation in yeast The Journal of biological chemistry High 17046832
2010 The N-terminal serine/threonine-rich domain (NSD) of yeast Mcm4 has both inhibitory and facilitating roles in DNA replication. The sole essential function of Dbf4-Cdc7 kinase (DDK) is to relieve the inhibitory activity within the NSD. Combining an mcm4 mutant lacking the NSD inhibitory domain with CDK-bypass mutations allows DNA synthesis in G1 phase. DDK is additionally required for intra-S checkpoint activation in response to hydroxyurea. Genetic epistasis in S. cerevisiae; mcm4 NSD deletion/mutation analysis; CDK-bypass mutations; whole-genome replication profiles; DNA damage checkpoint assays Nature High 20054399
2000 Electron microscopy reveals that the MCM4/6/7 complex forms ring-shaped toroidal structures with a central channel, consistent with a ring helicase. The complex binds single-stranded DNA at a level comparable to SV40 T antigen. MCM2 inhibits single-stranded DNA binding of MCM4/6/7, and the MCM3/5 complex inhibits its helicase activity. Electron microscopy with negative staining; gel-shift ssDNA binding assay; helicase inhibition assay with MCM2 or MCM3/5 Journal of molecular biology High 10884341
2002 Conserved ATPase motifs of MCM7 are essential for both ATPase and DNA helicase activities of the MCM4/6/7 complex. MCM7 requires interaction with other subunits for activity. The zinc finger of MCM4 is involved in subunit interactions between trimers. The N-terminal 35 or 112 amino acids of MCM4 are required for helicase activity of the hexameric complex even though complex formation is preserved. Baculovirus expression of mutant Mcm7 and Mcm4; ATPase and helicase assays; sedimentation analysis of complex formation The Journal of biological chemistry High 12207017
2003 The helicase and ATPase activities of mammalian MCM4/6/7 are specifically activated by thymine-rich single-stranded DNA sequences and by synthetic bubble or Y-fork structures that mimic activated replication origins, provided a sufficient-length thymine-containing ssDNA region is present. Sequences from the human lamin B2 replication origin activate the helicase; substitution of thymine clusters with guanine abolishes activation. In vitro ATPase and helicase assays with synthetic bubble/Y-fork substrates; systematic sequence variant analysis The EMBO journal High 14609960
1999 In budding yeast, CDKs (both G1-phase and B-type cyclins) cause nuclear export/exclusion of Mcm4 from the nucleus, providing a mechanism to prevent assembly of prereplicative complexes and block re-replication. B-type cyclins are required for origin firing, while G1 cyclins act to reduce nuclear Mcm4 before this. Live-cell imaging and immunofluorescence of GFP-tagged Mcm4 in yeast expressing different cyclin alleles; cell cycle synchronization Nature cell biology High 10559985
2000 In fission yeast, chromatin binding of mcm4/cdc21 occurs during anaphase B (earlier than in budding yeast) and requires orc1 and cdc18 (Cdc6 homolog). Release from chromatin occurs during S phase and requires ongoing DNA replication. Overexpression of cdc18 drives re-replication that depends on mcm4. GFP-tagging and detergent-wash chromatin-binding assay in S. pombe; genetic analysis with orc1 and cdc18 mutants; re-replication assay The EMBO journal High 10747035
2000 Distinct phosphorylation states of Xenopus Mcm4 regulate pre-replication complex assembly: an intermediately phosphorylated form is present in the transient interphase complex that loads onto chromatin; complete dephosphorylation inactivates Mcm complex and prevents chromatin binding; mitotic hyperphosphorylation requires Cdc2-cyclin B. Once on chromatin, Mcm4 and Mcm2 are the only subunits phosphorylated during pre-RC activation. Xenopus egg extract; kinase inhibitor treatments; CDK immunodepletion; chromatin fractionation; phosphorylation state analysis Molecular and cellular biology High 10779356
2006 During Epstein-Barr virus lytic replication, MCM4 is phosphorylated at Thr-19 and Thr-110 by CDK2/CDK1 and by EBV-encoded protein kinase (EBV-PK), inactivating the MCM4-6-7 helicase. EBV-PK expression in HeLa cells causes phosphorylation of these MCM4 sites and cell growth arrest. EBV-PK can also phosphorylate MCM6 and additional MCM4 sites beyond the CDK targets. In vitro kinase assay with purified MCM4-6-7; phospho-site mutagenesis; helicase activity assay; transfection of EBV-PK into HeLa cells Journal of virology High 17005684
2006 Site-specific phosphorylation of human MCM4 during the mammalian cell cycle: Thr7, Thr19, Ser32, Ser54, Ser88, and Thr110 are enhanced in G2/M phase (requiring CDK1), while Ser3 is phosphorylated during interphase (requiring CDK2). Different phosphorylation states result in different chromatin affinities; Ser32-phosphorylated MCM4 is enriched in the nucleolus throughout the cell cycle. Anti-phospho-MCM4 sera for seven individual sites; temperature-sensitive CDK1 mutant mouse cells; dominant-negative CDK2; chromatin fractionation; immunofluorescence The FEBS journal High 16519687
2008 The C-terminal domain (CTD) of Mcm4 in fission yeast is required to suspend MCM helicase activity after replication fork stalling, preventing excessive accumulation of single-stranded DNA. Truncation of the Mcm4 CTD (mcm4-c84) increases RPA association at stalled forks without affecting checkpoint activation or MCM/GINS association, indicating the CTD limits unwinding beyond that needed for checkpoint signaling. Fission yeast mcm4 CTD truncation mutant; 2D gel electrophoresis of replication intermediates; chromatin immunoprecipitation of RPA, MCM, GINS; HU sensitivity assays Proceedings of the National Academy of Sciences of the United States of America High 18753627
2014 The N-terminal disordered regulatory domain (NSD) of yeast Mcm4 integrates multiple kinase signals. The proximal NSD segment mediates checkpoint repression of late origins (Mec1/ATR pathway); the distal NSD segment controls replication fork progression and checkpoint signaling via CDK phosphorylation. These two sub-domains regulate origin timing and fork progression independently. Whole-genome replication profiling; single-molecule DNA fiber analysis; Mcm4 NSD segment-specific mutations; checkpoint kinase mutant analysis in S. cerevisiae Proceedings of the National Academy of Sciences of the United States of America High 24740181
2019 A conserved motif in Mcm4 is required for Mcm2-7 double-hexamer formation and extensive origin DNA unwinding. Mutations permitting loading of two Mcm2-7 complexes but blocking stable double-hexamer formation allow initial DNA melting and helicase-activation protein recruitment (Cdc45, GINS, Mcm10), but prevent extensive unwinding, identifying double-hexamer formation as the transition point. S. cerevisiae genetics; single-molecule imaging of MCM loading; helicase activation assays; biochemical reconstitution of MCM loading eLife High 31385807
2025 MCM2-7 ring closure at the Mcm2/Mcm5 interface triggers Mcm4 ATP hydrolysis, which drives reorganization of the MCM2-7 complex and Cdt1 release during pre-replicative complex assembly. The Mcm5 C-terminus contacts Orc3 and specifically recognizes the closed-ring conformation. Defective ring closure leads to MCM2-7 splitting and complex disassembly, identifying Mcm4 as the key ATPase regulating pre-RC formation. Cryo-EM structure of ORC-Cdc6-Cdt1-MCM2-7 intermediate; ATPase mutant analysis; Mcm2/Mcm5 interface mutagenesis; biochemical reconstitution Nature communications High 39747125
2008 S-CDK phosphorylation of five N-terminal CDK sites in yeast Mcm4 counteracts negative effects of ectopic DDK activation. Loss of these CDK sites (mcm4-5A) combined with DDK gain-of-function (mcm5/bob1 or DDK overexpression) is lethal, and deletion of S-phase cyclins (Clb5,6) is synthetically lethal with mcm4-5A, indicating that S-CDK and DDK cooperate through the Mcm4 N-terminal domain. S. cerevisiae genetics; CDK site alanine substitution; synthetic lethality assays; cyclin deletion analysis Molecular biology of the cell High 18321994
2010 ATP binding by MCM4/6/7 shifts the complex equilibrium from trimers toward hexamers; the minimal functional unit is a hexamer. Arginine finger mutations that disrupt inter-subunit ATP sites abolish helicase activity, indicating that all ATP sites formed in the hexameric configuration are required for full activity. Reconstitution of S. cerevisiae Mcm4/6/7 from individual subunits; sedimentation analysis; arginine finger mutagenesis; helicase and ATPase assays BMC biochemistry High 20860810
1995 Human MCM4 (hCdc21) forms a stable trimeric complex with two other human MCM proteins (p85Mcm and p105Mcm). MCM2/BM28 is more loosely associated. Phosphorylation of hCdc21 in mitotic cells correlates with reduced binding to nuclear structures. Co-immunoprecipitation; SDS-PAGE; immunofluorescence; cell cycle synchronization European journal of biochemistry Medium 7601140
1996 Xenopus cdc21 (MCM4 ortholog) binds to chromatin in an underphosphorylated state at the end of mitosis, is partially phosphorylated in early S phase before being displaced from chromatin, and is hyperphosphorylated by cdc2/cyclin B at the beginning of mitosis. Chromatin binding occurs prior to replication and is displaced as forks progress. Xenopus egg extracts; chromatin fractionation; indirect immunofluorescence; phosphorylation state analysis The EMBO journal High 8605878
1996 Human MCM4 (hCdc21), hCdc46, and P1Mcm3 are present on chromatin throughout the nucleus in late G1 and early S phase, but are displaced from chromatin specifically at sites of ongoing DNA replication during S phase, and are absent from chromatin in G2 and mitosis. They do not co-localize with replication foci but instead mark unreplicated chromatin. High-resolution confocal microscopy; immunofluorescence; comparison of MCM localization with BrdU-labeled replication foci in HeLa cells Journal of cell science High 8838654
2015 G364R mutation of MCM4 detected in human skin cancer cells reduces DNA helicase activity of the MCM4/6/7 complex to 30–50% of wild type without affecting complex formation, ssDNA binding, or ATPase activity. This mutant MCM4 localizes to chromatin-associated fraction in HeLa cells similarly to wild-type MCM4. In vitro expression and purification of mutant MCM4/6/7; helicase, ssDNA binding, and ATPase assays; immunofluorescence and chromatin fractionation in HeLa cells Journal of biochemistry High 25661590
2004 Levels of MCM4 phosphorylation inversely correlate with DNA synthesis levels during checkpoint control. Hydroxyurea or UV irradiation stimulates phosphorylation at CDK sites of MCM4, and this phosphorylation leads to inhibition of MCM4/6/7 helicase activity consistent with a role in suppressing DNA replication during checkpoint arrest. Phospho-specific antibody detection; [3H]-thymidine incorporation assays in HeLa cells treated with HU or UV Journal of structural biology Medium 15037254
2012 A Chaos3 mutation (F345I) in mouse Mcm4 destabilizes the MCM complex. Hypomorphic Mcm4 function causes chromosomal instability, enhanced sensitivity to replication inhibitor aphidicolin, and development of mammary adenocarcinomas in >80% of homozygous females. The corresponding yeast allele (F391I) causes minichromosome loss. Null Mcm4 causes preimplantation lethality. Mouse forward genetic screen; engineered yeast Mcm4 allele; chromosomal breakage assay in MEFs; tumor pathology Nature genetics High 17143284
2012 Partial MCM4 deficiency in humans (hypomorphic splice-site mutation producing loss of the major 96-kDa isoform while preserving the minor 85-kDa isoform) causes genomic instability in fibroblasts, rescuable by WT MCM4 expression. NK CD56(dim) cell deficiency results from impaired proliferation of NK CD56(bright) precursors, tightly dependent on MCM4-dependent cell division. Patient fibroblast genomic instability assay; WT MCM4 complementation; NK cell differentiation and proliferation assays; linkage and exome sequencing The Journal of clinical investigation High 22354167
2016 Concerted actions of Mcm4 NSD mutations, Sld3 mutations, and Dbf4 mutations are required to permit late origin firing under genotoxic stress (hydroxyurea). Checkpoint control of Sld3 impacts replication fork progression in parallel with Mcm4 NSD function; hypomorphic sld3 mutations are suppressed by mcm4 NSD mutations. S. cerevisiae genetics; whole-genome DNA replication profiling; hydroxyurea and dNTP limitation conditions; double and triple mutant analysis Genome research High 26733669
2012 The Mcm4(D573H) mutation (Sdl) in mice produces a dominant-negative, biologically inactive helicase when incorporated into MCM complexes. Sdl heterozygous mice develop T-ALL with high penetrance; homozygotes die during embryogenesis. Normal MCM levels are maintained, and the oncogenic effect results from dominant inactivation of MCM complexes rather than haploinsufficiency. Exome sequencing; S. cerevisiae helicase activity reconstitution; tumor pathology; micronucleated reticulocyte frequency; LOH analysis PLoS genetics High 23133403
2017 A G486D mutation in human MCM4 detected in endometrial cancer cells (within the conserved MCM-box) destabilizes the MCM4/6/7 complex by disrupting MCM4 interaction with MCM7, leading to degradation of mutant MCM4 protein. Expression of G486D MCM4 in HeLa cells induces abnormal nuclear morphology indicative of perturbed DNA replication. Baculovirus expression; co-immunoprecipitation; western blot; immunofluorescence in HeLa cells Journal of biochemistry Medium 27794528
2018 The N-terminal amino acids 1-35 of human MCM4, specifically Arg10 and Arg11, are required for DNA helicase activity of the MCM4/6/7 complex. CDK phospho-mimetic substitutions at all six N-terminal CDK sites reduce hexamer formation of MCM4/6/7, consistent with phosphorylation destabilizing the complex as a licensing control mechanism. Recombinant human MCM4 deletion and point mutants expressed in insect cells; helicase assays; size exclusion chromatography of MCM4/6/7 complex Journal of biochemistry Medium 30184107
2011 Heliquinomycin inhibits MCM4/6/7 DNA helicase activity (IC50 ~2.5 µM) by binding to single-stranded DNA and interfering with the ssDNA binding activity of the MCM4/6/7 complex. Treatment does not affect chromatin-bound MCM4 levels or activate the DNA replication stress checkpoint. Helicase activity assays with purified MCM4/6/7; gel retardation assay for ssDNA binding; chromatin fractionation in drug-treated cells; checkpoint marker analysis Journal of biochemistry Medium 22023799
2012 The mouse Chaos3 Mcm4 point mutation (F345I equivalent in human MCM4) perturbs MCM4 interaction with MCM6, reducing the formation of hexameric MCM4/6/7 complexes. This results in decreased chromatin binding of the MCM2-7 complex and abnormal nuclear localization of Mcm4 when expressed in human cells. Baculovirus co-expression; co-immunoprecipitation; size exclusion; immunofluorescence in human cells Journal of biochemistry Medium 22668557
1992 S. pombe cdc21 (MCM4 ortholog) is required for efficient DNA replication origin function, as shown by failure to maintain certain plasmids (minichromosome loss) and arrest with a nucleus containing two genome equivalents of DNA, indicating initiation or completion of replication at a subset of origins may fail. S. pombe genetic analysis; plasmid maintenance assay; FACS DNA content analysis Nucleic acids research Medium 1454522
1996 Fission yeast cdc21 is required for entry into S phase. Unlike budding yeast MCM proteins, cdc21 remains in the nucleus after S phase, indicating that nuclear export is not an essential regulatory mechanism in fission yeast. Depletion of cdc21 allows some cells to enter mitosis in the absence of DNA replication, suggesting cdc21 is needed for coordination between S phase and M phase. S. pombe gene deletion; DAPI/flow cytometry DNA content analysis; indirect immunofluorescence; nuclear localization assay The EMBO journal Medium 8631307
2005 On bubble substrates, MCM4/6/7 makes symmetric dual contacts with 5'-proximal 25 nt ssDNA segments at branch points, forming double hexamers for concurrent bidirectional unwinding. Loss of thymine residues from one strand significantly decreases unwinding efficiency, and the helicase is progressively inhibited by increasing GC content of the duplex region. In vitro helicase assays with defined synthetic DNA substrates (bubble, fork, extension); sequence substitution variants Nucleic acids research Medium 15917436
2004 Under-phosphorylated pRb (retinoblastoma protein) physically interacts with MCM4, as identified by yeast two-hybrid assay and confirmed by immunoprecipitation in Nalm-6 cells. A pRb-MCM4-CTF/NF-I complex was detected in cells. MCM4 contains a C-terminal motif with homology to the DNA-binding/viral replication activation domain of CTF/NF-I. LexA yeast two-hybrid assay; co-immunoprecipitation of pRb-MCM4-CTF complex from Nalm-6 cells; sequence homology analysis Biochemical and biophysical research communications Low 15081408

Source papers

Stage 0 corpus · 85 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1997 A DNA helicase activity is associated with an MCM4, -6, and -7 protein complex. The Journal of biological chemistry 461 9305914
2006 A viable allele of Mcm4 causes chromosome instability and mammary adenocarcinomas in mice. Nature genetics 257 17143284
2010 The Dbf4-Cdc7 kinase promotes S phase by alleviating an inhibitory activity in Mcm4. Nature 248 20054399
2012 Partial MCM4 deficiency in patients with growth retardation, adrenal insufficiency, and natural killer cell deficiency. The Journal of clinical investigation 224 22354167
2012 MCM4 mutation causes adrenal failure, short stature, and natural killer cell deficiency in humans. The Journal of clinical investigation 190 22354170
1999 Biochemical analysis of the intrinsic Mcm4-Mcm6-mcm7 DNA helicase activity. Molecular and cellular biology 180 10567526
1999 G1-phase and B-type cyclins exclude the DNA-replication factor Mcm4 from the nucleus. Nature cell biology 172 10559985
2006 Phosphorylation of MCM4 by Cdc7 kinase facilitates its interaction with Cdc45 on the chromatin. The Journal of biological chemistry 154 17046832
1996 Human replication proteins hCdc21, hCdc46 and P1Mcm3 bind chromatin uniformly before S-phase and are displaced locally during DNA replication. Journal of cell science 137 8838654
2003 Mcm4,6,7 uses a "pump in ring" mechanism to unwind DNA by steric exclusion and actively translocate along a duplex. The Journal of biological chemistry 135 13679365
1992 Fission yeast cdc21+ belongs to a family of proteins involved in an early step of chromosome replication. Nucleic acids research 117 1454522
1996 Phosphorylation of MCM4 by cdc2 protein kinase inhibits the activity of the minichromosome maintenance complex. Proceedings of the National Academy of Sciences of the United States of America 116 8901561
1996 Chromatin binding, nuclear localization and phosphorylation of Xenopus cdc21 are cell-cycle dependent and associated with the control of initiation of DNA replication. The EMBO journal 102 8605878
2014 Juvenile hormone-receptor complex acts on mcm4 and mcm7 to promote polyploidy and vitellogenesis in the migratory locust. PLoS genetics 94 25340846
2000 Chromatin binding of the fission yeast replication factor mcm4 occurs during anaphase and requires ORC and cdc18. The EMBO journal 90 10747035
2003 Identification of MCM4 as a target of the DNA replication block checkpoint system. The Journal of biological chemistry 87 12714602
1995 A human homologue of the yeast replication protein Cdc21. Interactions with other Mcm proteins. European journal of biochemistry 86 7601140
1987 The CDC8 transcript is cell cycle regulated in yeast and is expressed coordinately with CDC9 and CDC21 at a point preceding histone transcription. Experimental cell research 84 3305044
1996 Fission yeast cdc21, a member of the MCM protein family, is required for onset of S phase and is located in the nucleus throughout the cell cycle. The EMBO journal 83 8631307
2000 Inhibition of Mcm4,6,7 helicase activity by phosphorylation with cyclin A/Cdk2. The Journal of biological chemistry 80 10748114
2015 Proteome-wide analysis of mutant p53 targets in breast cancer identifies new levels of gain-of-function that influence PARP, PCNA, and MCM4. Proceedings of the National Academy of Sciences of the United States of America 79 25733866
2000 Electron microscopic observation and single-stranded DNA binding activity of the Mcm4,6,7 complex. Journal of molecular biology 78 10884341
2001 Phosphorylation of Mcm4 at specific sites by cyclin-dependent kinase leads to loss of Mcm4,6,7 helicase activity. The Journal of biological chemistry 76 11454864
1995 Molecular cloning of cDNA encoding mouse Cdc21 and CDC46 homologs and characterization of the products: physical interaction between P1(MCM3) and CDC46 proteins. Nucleic acids research 72 7610039
2016 MCM4 and MCM7, potential novel proliferation markers, significantly correlated with Ki-67, Bmi1, and cyclin E expression in esophageal adenocarcinoma, squamous cell carcinoma, and precancerous lesions. Human pathology 64 27476776
2002 Roles of Mcm7 and Mcm4 subunits in the DNA helicase activity of the mouse Mcm4/6/7 complex. The Journal of biological chemistry 64 12207017
1976 Yeast cell-cycle mutant cdc21 is a temperature-sensitive thymidylate auxotroph. Molecular & general genetics : MGG 62 794696
2006 Phosphorylation of MCM4 at sites inactivating DNA helicase activity of the MCM4-MCM6-MCM7 complex during Epstein-Barr virus productive replication. Journal of virology 54 17005684
2014 Domain within the helicase subunit Mcm4 integrates multiple kinase signals to control DNA replication initiation and fork progression. Proceedings of the National Academy of Sciences of the United States of America 47 24740181
2003 Thymine-rich single-stranded DNA activates Mcm4/6/7 helicase on Y-fork and bubble-like substrates. The EMBO journal 45 14609960
2021 Celastrol Modulates Multiple Signaling Pathways to Inhibit Proliferation of Pancreatic Cancer via DDIT3 and ATF3 Up-Regulation and RRM2 and MCM4 Down-Regulation. OncoTargets and therapy 44 34194230
2012 Recessive mutations in MCM4/PRKDC cause a novel syndrome involving a primary immunodeficiency and a disorder of DNA repair. Journal of medical genetics 42 22499342
2000 Distinct phosphoisoforms of the Xenopus Mcm4 protein regulate the function of the Mcm complex. Molecular and cellular biology 41 10779356
1998 The promoters for human DNA-PKcs (PRKDC) and MCM4: divergently transcribed genes located at chromosome 8 band q11. Genomics 40 9465298
2020 Antitumor Activity of Ohmyungsamycin A through the Regulation of the Skp2-p27 Axis and MCM4 in Human Colorectal Cancer Cells. Journal of natural products 39 31894983
2006 Site-specific phosphorylation of MCM4 during the cell cycle in mammalian cells. The FEBS journal 38 16519687
1995 Cdc54 belongs to the Cdc46/Mcm3 family of proteins which are essential for initiation of eukaryotic DNA replication. Gene 38 7698653
2012 A dominantly acting murine allele of Mcm4 causes chromosomal abnormalities and promotes tumorigenesis. PLoS genetics 34 23133403
2005 MCM4 expression in esophageal cancer from southern China and its clinical significance. Journal of cancer research and clinical oncology 34 16133572
2019 A conserved Mcm4 motif is required for Mcm2-7 double-hexamer formation and origin DNA unwinding. eLife 33 31385807
2016 Concerted activities of Mcm4, Sld3, and Dbf4 in control of origin activation and DNA replication fork progression. Genome research 27 26733669
2008 Interplay between S-cyclin-dependent kinase and Dbf4-dependent kinase in controlling DNA replication through phosphorylation of yeast Mcm4 N-terminal domain. Molecular biology of the cell 25 18321994
2008 Mcm4 C-terminal domain of MCM helicase prevents excessive formation of single-stranded DNA at stalled replication forks. Proceedings of the National Academy of Sciences of the United States of America 25 18753627
2004 Levels of MCM4 phosphorylation and DNA synthesis in DNA replication block checkpoint control. Journal of structural biology 25 15037254
2011 Widdrol activates DNA damage checkpoint through the signaling Chk2-p53-Cdc25A-p21-MCM4 pathway in HT29 cells. Molecular and cellular biochemistry 23 22160829
2010 C. elegans MCM-4 is a general DNA replication and checkpoint component with an epidermis-specific requirement for growth and viability. Developmental biology 23 21146520
2007 Genetic screen for chromosome instability in mice: Mcm4 and breast cancer. Cell cycle (Georgetown, Tex.) 22 17495541
2005 DNA binding and helicase actions of mouse MCM4/6/7 helicase. Nucleic acids research 20 15917436
2015 G364R mutation of MCM4 detected in human skin cancer cells affects DNA helicase activity of MCM4/6/7 complex. Journal of biochemistry 19 25661590
2010 Increase of Mcm3 and Mcm4 expression in cervical squamous cell carcinomas. European journal of gynaecological oncology 19 21077471
2012 Effect of an MCM4 mutation that causes tumours in mouse on human MCM4/6/7 complex formation. Journal of biochemistry 16 22668557
2015 High expression of carbonic anhydrase IX is significantly associated with glandular lesions in gastroesophageal junction and with tumorigenesis markers BMI1, MCM4 and MCM7. BMC gastroenterology 15 26156831
2012 The Thermococcus kodakaraensis Tko CDC21-1 intein activates its N-terminal splice junction in the absence of a conserved histidine by a compensatory mechanism. Biochemistry 15 22380677
2011 Characterization of Leishmania donovani MCM4: expression patterns and interaction with PCNA. PloS one 15 21829589
2017 An MCM4 mutation detected in cancer cells affects MCM4/6/7 complex formation. Journal of biochemistry 14 27794528
2011 Interaction of heliquinomycin with single-stranded DNA inhibits MCM4/6/7 helicase. Journal of biochemistry 13 22023799
2024 MCM4 potentiates evasion of hepatocellular carcinoma from sorafenib-induced ferroptosis through Nrf2 signaling pathway. International immunopharmacology 12 39276458
2015 HPV-type-specific response of cervical cancer cells to cisplatin after silencing replication licensing factor MCM4. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 12 26188903
2023 EZH2 regulates pancreatic cancer cells through E2F1, GLI1, CDK3, and Mcm4. Hereditas 11 37198697
2001 Identification of an MCM4 homologue expressed specifically in the sexual stage of Plasmodium falciparum. International journal for parasitology 9 11513894
1997 MCM4 and PRKDC, human genes encoding proteins MCM4 and DNA-PKcs, are close neighbours located on chromosome 8q12-->q13. Cytogenetics and cell genetics 9 9284934
2019 Crystal structures of CDC21-1 inteins from hyperthermophilic archaea reveal the selection mechanism for the highly conserved homing endonuclease insertion site. Extremophiles : life under extreme conditions 8 31363851
2025 MCM2-7 ring closure involves the Mcm5 C-terminus and triggers Mcm4 ATP hydrolysis. Nature communications 7 39747125
2024 Identification of MCM4 and PRKDC as new regulators of osteosarcoma cell dormancy based on 3D cell cultures. Biochimica et biophysica acta. Molecular cell research 7 38216092
2021 MCM4 Is a Novel Biomarker Associated With Genomic Instability, BRCAness Phenotype, and Therapeutic Potentials in Soft-Tissue Sarcoma. Frontiers in cell and developmental biology 7 34178990
2018 Function of the amino-terminal region of human MCM4 in helicase activity. Journal of biochemistry 7 30184107
2016 Characterization of a Novel MMS-Sensitive Allele of Schizosaccharomyces pombe mcm4. G3 (Bethesda, Md.) 6 27473316
2010 The effects of oligomerization on Saccharomyces cerevisiae Mcm4/6/7 function. BMC biochemistry 6 20860810
2020 Effect of the natural compound trans‑resveratrol on human MCM4 gene transcription. Oncology reports 5 32377740
2015 c-ETS transcription factors play an essential role in the licensing of human MCM4 origin of replication. Biochimica et biophysica acta 5 26365772
2007 [Expression and significance of MCM4 in esophageal cancer]. Ai zheng = Aizheng = Chinese journal of cancer 5 17222376
2004 MCM4 shares homology to a replication/DNA-binding domain in CTF and is contacted by pRb. Biochemical and biophysical research communications 5 15081408
2025 Celastrol promotes DNA damage and apoptosis in uterine corpus endometrial carcinoma via promotion of KAT2B-mediated RBPJ acetylation and repression of MCM4 transcription. Molecular medicine (Cambridge, Mass.) 4 39901144
2020 Active Replication Checkpoint Drives Genome Instability in Fission Yeast mcm4 Mutant. Molecular and cellular biology 4 32341083
2024 MCM4 Promotes the Progression of Malignant Melanoma by Activating the PI3K/AKT Pathway. Environmental toxicology 3 39501995
2023 E2F2 is upregulated by the ERK pathway and regulates decidualization via MCM4. Gene 2 37028609
2013 Expression, purification and biochemical characterization of Schizosaccharomyces pombe Mcm4, 6 and 7. BMC biochemistry 2 23444842
2007 The promoter regions of the Myb-regulated Adora2B and Mcm4 genes co-localize with origins of DNA replication. BMC molecular biology 2 17822556
2025 E2F7 upregulates MCM4 and fatty acid metabolism to advance lung adenocarcinoma metastasis. Prostaglandins & other lipid mediators 1 40158794
2025 Transcription factor E2F1 promotes non-small cell lung cancer progression by activating the PI3K/AKT pathway through MCM4. Journal of cardiothoracic surgery 1 40437556
2025 MCM4 as Potential Metastatic Biomarker in Lung Adenocarcinoma. Diagnostics (Basel, Switzerland) 1 40564876
2026 Expression of MCM2, MCM4, and MCM10 in hepatocellular carcinoma based on bioinformatic analyses and their predictive value for postoperative recurrence: An initial model development study. Surgical oncology 0 41921256
2025 Promotion of Melanoma Progression through MCM4-Induced Immune Suppression and Polarization of Macrophages by Carcinogenic Exosomes. Current cancer drug targets 0 40375706
2025 Comprehensive analysis reveals MCM4 as a biomarker for guiding therapies and immunomodulatory role in skin cutaneous melanoma. Journal of Cancer 0 40959096
2025 Integrative Multi-Omics and Functional Characterization Reveal MCM4 as a Key Oncogenic Regulator in Hepatocellular Carcinoma. OncoTargets and therapy 0 41333157