Affinage

CDT1

DNA replication factor Cdt1 · UniProt Q9H211

Length
546 aa
Mass
60.4 kDa
Annotated
2026-04-28
100 papers in source corpus 47 papers cited in narrative 47 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CDT1 is a conserved replication licensing factor that loads the MCM2-7 helicase onto replication origins during G1 phase to ensure DNA replicates exactly once per cell cycle. CDT1 cooperates with Cdc6/ORC to form pre-replicative complexes, physically embracing Mcm2, Mcm4, and Mcm6 within an open-ring heptamer that is deposited onto origin DNA, and it directly stimulates MCM helicase activity in vitro while inhibiting CMG helicase at early replication forks until it is degraded (PMID:10766248, PMID:28191893, PMID:18606811, PMID:36608667). CDT1 abundance is tightly controlled by multiple overlapping ubiquitin-dependent pathways—CRL4(DDB1/CDT2) acting via a PCNA-bound PIP-box in S phase and after DNA damage, SCF-Skp2 recognizing CDK-phosphorylated CDT1, APC/C(Cdh1) via N-terminal destruction boxes, and SCF-FBXO31 in G2—with p97/UFD1 extracting ubiquitinated CDT1 from chromatin for proteasomal destruction (PMID:16362051, PMID:16482215, PMID:18162579, PMID:24828503, PMID:21981919). The inhibitor geminin forms a stoichiometry-dependent heterohexamer with CDT1 that sterically blocks MCM access while paradoxically stabilizing CDT1 protein during S phase and mitosis, and additional modulators including FOXO3, USP37, and KAT2B/KAT3B-mediated acetylation further tune CDT1 levels to enforce the once-per-cell-cycle replication rule (PMID:19906994, PMID:15257290, PMID:22451935, PMID:27296872, PMID:19276081).

Mechanistic history

Synthesis pass · year-by-year structured walk · 19 steps
  1. 1994 High

    Identification of CDT1 as an essential S-phase factor established that a previously unknown gene product was required for DNA replication initiation but not for mitotic entry, opening the question of how it acts mechanistically.

    Evidence Genetic screen and null allele analysis in fission yeast

    PMID:8313888

    Open questions at the time
    • Biochemical function unknown
    • No physical interactions identified
    • No connection to licensing machinery
  2. 2000 High

    Demonstration that Cdt1 cooperates with Cdc6 to load MCM onto chromatin and that geminin directly inhibits Cdt1 placed CDT1 at the heart of the replication licensing mechanism and identified the first negative regulator.

    Evidence Chromatin fractionation and re-replication assays in fission yeast; cell-free Xenopus replication with geminin inhibition/rescue

    PMID:10766248 PMID:11125146

    Open questions at the time
    • How CDT1 physically contacts MCM or ORC unknown
    • Mechanism of geminin inhibition unresolved
    • Post-translational regulation of CDT1 unexplored
  3. 2002 High

    Mapping of CDT1's direct binding partners (Mcm6, ORC2, geminin) and its intrinsic DNA-binding activity defined the molecular contacts through which CDT1 bridges origin recognition and helicase loading.

    Evidence Yeast two-hybrid, in vitro DNA-binding, and domain mapping with purified mouse Cdt1; human CDT1 antibody microinjection blocking replication and MCM loading

    PMID:11836525 PMID:11896191 PMID:12192004

    Open questions at the time
    • Structural basis of CDT1-MCM and CDT1-geminin interactions unknown
    • Mechanism of CDT1 turnover after S phase entry unexplored
  4. 2003 High

    Discovery that SCF-Skp2 ubiquitinates CDT1 in a CDK-phosphorylation-dependent manner and that CUL4-ROC1 degrades CDT1 after DNA damage revealed the first two E3 ligase pathways controlling CDT1 abundance, framing the concept of multi-layered proteolytic control.

    Evidence In vitro and in vivo ubiquitination assays with Skp2; siRNA against CUL4-ROC1 and DNA damage-induced stability assays

    PMID:12840033 PMID:14578910

    Open questions at the time
    • Substrate receptor for CUL4 pathway not yet identified
    • Relative contributions of each E3 ligase during normal S phase unclear
    • Role of PCNA not yet connected
  5. 2004 High

    The crystal structure of the geminin-Cdt1 complex revealed that geminin's coiled-coil dimer sterically blocks MCM access to Cdt1, while parallel work showed that geminin paradoxically stabilizes CDT1 protein during S/M phase by preventing its ubiquitination, resolving geminin's dual role.

    Evidence X-ray crystallography with mutagenesis; siRNA knockdown of geminin with ubiquitination and stability assays

    PMID:15257290 PMID:15286659

    Open questions at the time
    • Stoichiometric switch between permissive and inhibitory geminin-CDT1 complexes not yet characterized
    • Whether geminin protects CDT1 from specific E3 ligases unknown
  6. 2004 High

    Demonstration that CDK2/4-mediated phosphorylation of CDT1 at the Cy motif promotes Skp2 binding, and that CDT1 ubiquitination on chromatin requires replication initiation factors, established the coupling between S-phase entry and CDT1 destruction.

    Evidence In vitro kinase assays with phosphopeptide mapping and Cy-motif mutagenesis; Xenopus egg extract replication assays with factor depletions

    PMID:15004027 PMID:15598982

    Open questions at the time
    • PCNA involvement not yet directly demonstrated
    • Identity of the replication-coupled E3 ligase still unclear
  7. 2005 High

    Discovery that PCNA directly binds CDT1 through a PIP-box and is required for CRL4/DDB1-mediated CDT1 destruction unified the replication-coupled degradation pathway and explained how ongoing replication triggers CDT1 removal.

    Evidence Xenopus egg extract with PCNA depletion, PIP-box mutagenesis causing re-replication, and chromatin fractionation showing DDB1 recruitment depends on CDT1-PCNA

    PMID:15855168 PMID:16362051

    Open questions at the time
    • The specific WD40-repeat adaptor connecting DDB1 to CDT1 not yet identified
    • N-terminal degron versus PIP-box contributions not fully separated
  8. 2006 High

    Systematic dissection revealed that two distinct E3 ligases—CRL4(DDB1) and SCF-Skp2—target different CDT1 degrons with different cell-cycle timing, and that CDT2/DTL is the substrate adaptor bridging DDB1 to CDT1, completing the molecular identity of the primary S-phase degradation pathway.

    Evidence siRNA co-knockdown of both E3 pathways with mutagenesis; CDT2 co-IP with CUL4-DDB1 and genetic rescue in zebrafish

    PMID:16482215 PMID:16940174 PMID:17085480

    Open questions at the time
    • E2 enzymes for each pathway not identified
    • Whether additional adaptor proteins modulate the CRL4-CDT2 pathway unknown
  9. 2007 High

    Identification of APC/C(Cdh1) as a third E3 ligase targeting CDT1 via N-terminal destruction boxes, and of the Orc6-Cdt1 dynamic interaction required for repeated MCM loading, expanded the regulatory circuit and refined the loading mechanism.

    Evidence Proteomic identification of APC/C-CDT1 interaction with D-box mutagenesis causing re-replication; reconstituted in vitro MCM loading with Orc6 domain fusions

    PMID:18006685 PMID:18162579

    Open questions at the time
    • How APC/C(Cdh1) coordinates with other E3 ligases at M/G1 transition unclear
    • Structural basis of Cdt1-ORC6 interaction lacking
  10. 2008 High

    Demonstration that Cdt1 directly stimulates MCM4/6/7 helicase activity in vitro established a catalytic role for Cdt1 beyond passive recruitment, and identification of MCM9 as a CDT1 partner that links Cdt1 to MCM2-7 added a new layer to the loading pathway.

    Evidence In vitro helicase assays with purified proteins and CDT1 mutants; co-IP and immunodepletion of MCM9 in Xenopus egg extracts

    PMID:18606811 PMID:18657502

    Open questions at the time
    • Physiological relevance of CDT1 helicase stimulation during normal replication unknown
    • Whether MCM9-CDT1 interaction is conserved in all vertebrates not tested
  11. 2009 High

    Resolution of the geminin-CDT1 heterohexamer crystal structure revealed a stoichiometry-dependent molecular switch between a permissive heterotrimer and an inhibitory heterohexamer, while discovery of CDT1 acetylation by KAT2B/KAT3B (antagonized by HDAC11) identified a non-proteolytic regulatory axis.

    Evidence X-ray crystallography and SAXS of geminin-CDT1 with functional assays; in vitro acetylation assay and ubiquitination protection experiments

    PMID:19276081 PMID:19906994

    Open questions at the time
    • What controls the heterotrimer-to-heterohexamer transition in vivo unknown
    • Which specific lysines are acetylated and their individual contributions unresolved
  12. 2010 High

    NMR structures of the Mcm6 C-terminal winged-helix domain bound to CDT1 defined the atomic interface essential for MCM loading, and CDT1 was shown to induce HBO1-dependent chromatin decondensation at origins during G1.

    Evidence NMR structure with interface mutagenesis abolishing MCM loading and yeast viability; chromatin decondensation assays with HBO1 knockdown

    PMID:20202939 PMID:20980834

    Open questions at the time
    • Chromatin decondensation role observed in single lab with artificial arrays
    • Full-length CDT1-MCM6 complex structure not available
  13. 2011 High

    Multiple findings expanded understanding of CDT1 regulation: JNK1 phosphorylation of CDT1-T29 blocks HBO1-dependent origin firing under stress; CRL4(CDT2) uses UBE2G1/2 E2 enzymes specifically for CDT1 ubiquitination; p97/UFD1 extracts ubiquitinated CDT1 from chromatin; and CDT1 is rapidly recruited to DNA damage sites by PCNA.

    Evidence In vitro kinase assay with ChIP; proteomic and siRNA screens with in vitro ubiquitination; genome-wide screen validated in Xenopus extracts; laser microsurgery with live-cell imaging

    PMID:21224399 PMID:21628527 PMID:21856198 PMID:21981919

    Open questions at the time
    • Relative importance of JNK1 versus CDK phosphorylation at T29 in vivo unclear
    • p97 substrate selectivity for CDT1 versus other chromatin substrates not defined
  14. 2012 Medium

    FOXO3 was identified as a positive CDT1 regulator that forms a complex with CDT1 blocking DDB1/PCNA access, establishing that CDT1 stability is not solely determined by E3 ligases but also by protective protein-protein interactions.

    Evidence Co-immunoprecipitation and siRNA knockdown with cell cycle analysis

    PMID:22451935

    Open questions at the time
    • Single-lab observation awaits independent replication
    • Whether FOXO3-CDT1 interaction is regulated by FOXO3 phosphorylation not tested
    • Structural basis unknown
  15. 2014 Medium

    Discovery of SCF-FBXO31 as a fourth E3 ligase targeting CDT1 specifically in G2 phase completed the cell-cycle-phase-specific map of CDT1 destruction pathways.

    Evidence Co-immunoprecipitation, ubiquitination assay, cell cycle synchronization, re-replication assay

    PMID:24828503

    Open questions at the time
    • Single-lab observation
    • The degron recognized by FBXO31 not mapped
    • Relative contribution to CDT1 regulation versus other E3 pathways unknown
  16. 2016 Medium

    USP37 was identified as a deubiquitinase that directly stabilizes CDT1 during G1/S, affecting MCM loading and replication fork speed, adding a deubiquitination axis to CDT1 regulation.

    Evidence Co-immunoprecipitation, in vivo de-ubiquitination assay, DNA fiber assay, siRNA knockdown

    PMID:27296872

    Open questions at the time
    • Single-lab observation
    • Which ubiquitin chains USP37 removes from CDT1 not characterized
    • Interplay with geminin-mediated protection unknown
  17. 2017 High

    Cryo-EM structures of the OCCM loading intermediate at near-atomic resolution revealed that Cdt1 adopts a three-domain configuration wrapping around Mcm2/4/6, stabilizing the open MCM ring and positioning it for double-hexamer formation, providing the definitive structural framework for CDT1's role in helicase loading.

    Evidence Cryo-EM at 3.9 Å with cross-linking mass spectrometry of yeast OCCM complex

    PMID:28191893 PMID:28191894

    Open questions at the time
    • Human OCCM structure not available
    • Mechanism of CDT1 release after MCM loading not resolved structurally
    • No time-resolved structure of the loading process
  18. 2021 High

    SLFN11 was shown to promote CDT1 degradation by binding DDB1 at replication forks under damage conditions, revealing a damage-specific accessory factor for CRL4(CDT2) and explaining how SLFN11-expressing cells irreversibly block replication after damage.

    Evidence Co-immunoprecipitation, siRNA knockdown, domain mutagenesis, protein stability assays

    PMID:33536335

    Open questions at the time
    • Whether SLFN11 is required for CDT1 degradation in unperturbed S phase unknown
    • Structural basis of SLFN11-DDB1 interaction not determined
  19. 2023 High

    In vitro reconstitution with purified human proteins demonstrated that CDT1 inhibits CMG helicase activity at fired origins, and that CRL4(CDT2)-mediated CDT1 destruction is required not just to prevent re-licensing but to permit DNA synthesis, revealing a previously unrecognized fork-level function.

    Evidence Reconstituted human DNA replication system with purified proteins and quantitative single-cell microscopy

    PMID:36608667

    Open questions at the time
    • Mechanism by which CDT1 inhibits CMG at a structural level unknown
    • Whether CDT1's inhibitory role at forks contributes to checkpoint signaling unexplored
    • Temporal overlap between licensing and firing not quantified genome-wide

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key unresolved questions include: the structural mechanism by which CDT1 inhibits CMG helicase, how CDT1 is released from the MCM ring after double-hexamer formation, the full human OCCM structure, how the multiple E3 ligase and stabilizing pathways are coordinated in a single cell cycle, and whether CDT1's chromatin-remodeling activity is a general feature of origin licensing.
  • No structural model of CDT1-CMG inhibitory complex
  • Mechanism of CDT1 release after MCM loading unknown
  • Integration of four E3 ligase pathways and multiple stabilizing factors not modeled quantitatively

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0008092 cytoskeletal protein binding 4 GO:0098772 molecular function regulator activity 2 GO:0003677 DNA binding 1
Localization
GO:0005694 chromosome 4 GO:0005634 nucleus 3
Pathway
R-HSA-69306 DNA Replication 8 R-HSA-392499 Metabolism of proteins 6 R-HSA-1640170 Cell Cycle 5 R-HSA-73894 DNA Repair 5
Partners
CDC6DDB1GMNNHDAC11MCM2MCM6ORC6PCNA
Complex memberships
Cdt1-MCM9 complexGeminin-Cdt1 heterohexamerOCCM (ORC-Cdc6-Cdt1-Mcm2-7)pre-replicative complex (pre-RC)

Evidence

Reading pass · 47 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1994 CDT1 (cdt1) is a transcriptional target of the Cdc10/Sct1 transcription factor in fission yeast and is essential for DNA replication but not for mitotic initiation, placing it as a required factor for S phase that also maintains the normal S phase-mitosis dependency relationship. Genetic screen, immunoprecipitation-PCR, null allele analysis, ectopic expression rescue The EMBO journal High 8313888
2000 Fission yeast Cdt1 cooperates with Cdc18 (Cdc6) to load the MCM protein Cdc21 onto chromatin at the end of mitosis, is located in the nucleus, peaks in expression as cells finish mitosis, and interacts physically with Cdc18. Re-replication assay, chromatin fractionation, immunoprecipitation, cell cycle synchronization Nature High 10766248
2000 Geminin binds tightly to Cdt1 and inhibits DNA replication by targeting Cdt1; excess Cdt1 reverses geminin-mediated inhibition of DNA replication in cell-free Xenopus extracts. Cell-free DNA replication assay, binding studies, rescue with excess Cdt1 Science High 11125146
2001 Human CDT1 is a nuclear protein present only during G1 phase; after S phase onset its levels decrease via proteasome-mediated proteolysis (not transcriptional downregulation), and it can associate with geminin. Cell cycle synchronization, immunoblotting, proteasome inhibitor treatment, co-immunoprecipitation The Journal of biological chemistry High 11555648
2002 Budding yeast Cdt1 is required for pre-RC assembly; it accumulates in the nucleus during G1 and is excluded by CDKs in S/G2; Cdt1 interacts with the Mcm2-7 complex and their nuclear accumulations are interdependent. Genetic analysis, co-immunoprecipitation, cell fractionation, nuclear localization assays Nature cell biology High 11836525
2002 Human CDT1 is essential for DNA replication and for loading MCM proteins onto chromatin; antibody microinjection blocks DNA replication and MCM chromatin loading. Antibody microinjection, DNA replication assay, chromatin fractionation Journal of cell science High 11896191
2002 Mouse Cdt1 directly binds to Mcm6 (C-terminal region, aa 407-477), ORC2, and geminin (central region, aa 177-380) by yeast two-hybrid; Cdt1 has intrinsic sequence-independent DNA binding activity that is inhibited by geminin. Yeast two-hybrid, protein purification, in vitro DNA binding assay, domain mapping The Journal of biological chemistry High 12192004
2003 SCF-Skp2 ubiquitin ligase interacts with human CDT1 in a phosphorylation-dependent manner and ubiquitinates it in vivo and in vitro; Skp2 knockdown or disruption of Cdt1-Skp2 interaction stabilizes Cdt1. Co-immunoprecipitation, in vivo and in vitro ubiquitination assay, siRNA knockdown The Journal of biological chemistry High 12840033
2003 Cdc6 physically associates with CDT1 via its N-terminal noncatalytic domain in mammalian cells; Cdc6 induces binding of Mcm2 to Cdt1; geminin blocks Cdt1 interaction with both Mcm2 and Cdc6. Co-immunoprecipitation, ectopic expression, cell cycle analysis The Journal of biological chemistry High 14672932
2003 CDT1 is rapidly proteolysed after UV or gamma-irradiation by CUL4-ROC1 ubiquitin E3 ligase complexes; this proteolysis is independent of ATM/CHK2 and constitutes a novel G1 DNA damage checkpoint; loss of COP9-signalosome or CUL4-ROC1 suppresses CDT1 proteolysis. Protein stability assays, siRNA knockdown, ubiquitination assays, IR/UV treatment Nature cell biology High 14578910
2004 Crystal structure of the geminin-Cdt1 complex reveals that the amino-terminal coiled-coil region of geminin dimer interacts with both N-terminal and C-terminal parts of Cdt1, with key interface residues Pro181, Ala182, Tyr183, Phe186, and Leu189 of Cdt1; the C-terminal region of geminin sterically hinders MCM access to Cdt1. X-ray crystallography, biochemical binding assays, mutagenesis Nature High 15286659
2004 CDT1 is destroyed by ubiquitin-mediated proteolysis during the first round of DNA replication in Xenopus egg extracts in a manner requiring Cdk2/Cyclin E, Cdc45, RPA, and polymerase alpha (i.e., replication initiation); CDT1 is ubiquitinated on chromatin; this acts redundantly with geminin to prevent re-replication. Xenopus egg extract replication assay, immunodepletion, ubiquitination assay Genes & development High 15598982
2004 DDB1 directly binds CDT1 in vitro and bridges it to CUL4A in vivo; DDB1 silencing prevents UV-induced CDT1 degradation and CUL4A-mediated CDT1 ubiquitination, identifying DDB1 as the substrate receptor of a CUL4A(DDB1) E3 ligase for CDT1. In vitro direct binding, co-immunoprecipitation, siRNA knockdown, in vitro ubiquitination Nature cell biology High 15448697
2004 Human CDT1 is phosphorylated by Cdk2 and Cdk4 in vivo and in vitro; phosphorylation promotes binding to Skp2; a CDT1 mutant at the Cy motif/Cdk binding site is not phosphorylated, fails to bind Skp2, and is more stable. In vitro kinase assay, phosphopeptide mapping, co-immunoprecipitation, mutagenesis The Journal of biological chemistry High 15004027
2004 Geminin stabilizes CDT1 protein from proteasome-mediated degradation during S phase and mitosis by inhibiting its ubiquitination; this accumulation of CDT1 in mitosis is required for pre-RC formation and DNA replication in the following cell cycle. siRNA knockdown, proteasome inhibitor, ubiquitination assay, cell cycle analysis The EMBO journal High 15257290
2004 CDT1 is degraded after UV-induced DNA damage via the SCF-Skp2 complex in a phosphorylation- and caffeine-sensitive (checkpoint-dependent) manner; overexpression of CDT1 after UV interferes with inhibition of DNA synthesis. Protein stability assays, co-immunoprecipitation, proteasome inhibition, overexpression The Journal of biological chemistry Medium 15102855
2005 CDT1 binds PCNA through a conserved PIP-box motif; PCNA depletion from Xenopus egg extracts inhibits replication-dependent CDT1 destruction; mutation of the PCNA-interaction motif yields a stable CDT1 that induces re-replication; DDB1 chromatin loading depends on CDT1-PCNA binding, indicating PCNA activates the pre-formed CDT1-CUL4(DDB1) ligase complex. Xenopus egg extract assay, PCNA immunodepletion, PIP-box mutagenesis, re-replication assay, chromatin fractionation Nature cell biology High 16362051
2005 CDT1 degradation in S phase requires its N-terminal 32 residues via a Skp2-independent, PCNA-dependent pathway; Cdk2-phosphorylation at Thr29 promotes Skp2 binding but is not necessary for S-phase proteolysis; stabilized CDT1 delays S phase progression. Phosphopeptide mapping, mutagenesis, S-phase progression assay, co-immunoprecipitation The Journal of biological chemistry High 15855168
2006 CDT1 is targeted for proteolysis by two distinct E3 ubiquitin ligases: CUL4-DDB1 (recognizing six conserved N-terminal aa; active during S phase replication and after DNA damage; requires PCNA) and SCF-Skp2 (recognizing CDK-phosphorylated Cy-motif; active in S and G2); co-silencing both stabilizes CDT1 throughout S-G2. siRNA co-knockdown, mutagenesis, ubiquitination assays, cell cycle analysis The EMBO journal High 16482215
2006 PCNA is required for CUL4-DDB1-mediated CDT1 degradation after DNA damage in mammalian cells and fission yeast; PCNA inhibitory domain of p21/p57 blocks CDT1 degradation; DDB1 deletion in fission yeast also accumulates CDT1 in undamaged cells. siRNA knockdown, PCNA inhibitory domain overexpression, fission yeast genetics The Journal of biological chemistry High 16407242
2006 PCNA is a cofactor for CUL4/DDB1-mediated N-terminal ubiquitination of CDT1; the PCNA-binding motif and Cul4/Ddb1 are required for this S-phase and DNA damage-induced degradation pathway. Mutagenesis, ubiquitination assays, siRNA knockdown, co-immunoprecipitation The Journal of biological chemistry High 16407252
2006 DTL/CDT2 associates with CUL4-DDB1 E3 ubiquitin ligase and is required for CDT1 down-regulation in unperturbed S phase and following DNA damage; genetic rescue experiments show that CDT1 reduction suppresses rereplication defects of Dtl-deficient zebrafish. Co-immunoprecipitation, siRNA knockdown, zebrafish genetics, epistasis analysis Genes & development High 17085480
2006 DDB1 loss leads to CDT1 accumulation, rereplication, and DNA double-strand break accumulation with ATM/ATR checkpoint activation; CDT1 co-depletion partially suppresses these phenotypes, establishing CDT1 regulation by DDB1-CUL4A as critical for genome stability. siRNA knockdown, epistasis by co-depletion, flow cytometry, γH2AX assay Molecular and cellular biology High 16940174
2007 Orc6 directly binds Cdt1 through two regions; ORC lacking Orc6 fails to interact with Cdt1 or load Mcm2-7; a fusion of Cdt1 to Orc6-CTD restores single-round but not multiple-round Mcm2-7 loading, demonstrating that dynamic Cdt1-ORC6 association is required for repeated MCM loading. In vitro Mcm2-7 loading assay with purified proteins, direct binding studies, domain fusion experiments Genes & development High 18006685
2007 Nuclear accumulation of cyclin D1 during S phase stabilizes CDT1 by inhibiting CUL4-dependent CDT1 proteolysis, triggering DNA rereplication and p53-dependent apoptosis. Mouse models, protein stability assays, cell cycle analysis, re-replication assay Genes & development High 18006686
2007 APC/C(Cdh1) ubiquitin ligase controls CDT1 levels via three destruction boxes in the CDT1 N-terminus; elimination of these destruction boxes causes strong rereplication and chromosomal damage. Proteomics/affinity chromatography-MS to identify CDT1-binding proteins, co-immunoprecipitation, mutagenesis, re-replication assay Molecular biology of the cell High 18162579
2007 CDT1 dynamically associates with chromatin throughout G1; CDT1 simultaneously binds Geminin and chromatin in vivo, recruiting Geminin onto chromatin; the domains for chromatin binding and Geminin binding are separable. Quantitative live-cell fluorescence microscopy (FRAP), domain mapping, co-localization in living cells The EMBO journal High 17318181
2008 MCM9 forms a stable complex with CDT1 and is required for recruiting MCM2-7 helicase onto chromatin; MCM9 prevents excess geminin accumulation at chromatin during licensing and acts as an activating linker between Cdt1 and MCM2-7. Co-immunoprecipitation, chromatin fractionation, immunodepletion in Xenopus egg extracts Molecular cell High 18657502
2008 Cdt1 forms a complex with Mcm4/6/7 and Mcm2-7 and directly stimulates the DNA binding and helicase activities of Mcm4/6/7 in vitro; a Cdt1 mutant defective in MCM stimulation has reduced MCM interaction and fails to form a high molecular weight complex with MCM. Glycerol gradient fractionation, in vitro helicase assay, mutagenesis, protein binding assays The Journal of biological chemistry High 18606811
2009 CDT1 undergoes acetylation at its N-terminus by lysine acetyltransferases KAT2B and KAT3B; it is deacetylated by HDAC11 which directly binds CDT1; acetylation protects CDT1 from ubiquitylation and proteasomal degradation. Co-immunoprecipitation, in vitro acetylation assay, ubiquitination assay, proteasome inhibition The Journal of biological chemistry High 19276081
2009 The human CDT1-Geminin complex can exist as a permissive heterotrimer or an inhibitory heterohexamer; specific CDT1 residues buried in the heterohexamer are required for licensing; the transition between these forms represents a molecular switch between licensing-competent and licensing-defective states. X-ray crystallography, SAXS, functional assays in Xenopus and mammalian cells Proceedings of the National Academy of Sciences of the United States of America High 19906994
2010 CDT1 induces large-scale chromatin decondensation in G1 required for MCM loading; this requires HBO1 HAT activity and histone H4 modifications; HDAC11 bound to CDT1 in S phase suppresses chromatin unfolding and MCM loading; Geminin-HDAC11 interaction with CDT1 represses this activity. Chromatin decondensation assay, siRNA knockdown, MCM loading assay, co-immunoprecipitation Cell cycle (Georgetown, Tex.) Medium 20980834
2010 The C-terminal domain of human Mcm6 (Cdt1 binding domain, CBD) has a winged-helix fold and mediates specific interaction with CDT1 through its helix-turn-helix motif; NMR structure resolved; charge complementarity is key; mutations in this interface abolish MCM2-7 chromatin loading and yeast viability. NMR structure determination, mutagenesis, yeast genetics, chromatin loading assay The Journal of biological chemistry High 20202939
2011 JNK1 phosphorylates CDT1 on threonine 29 in response to non-genotoxic stress, causing dissociation of the HBO1 histone acetylase from replication origins and blocking initiation of DNA replication. In vitro kinase assay, chromatin immunoprecipitation, replication origin assay, stress response experiments Molecular cell High 21856198
2011 CRL4(CDT2) utilizes UBE2G family E2 ubiquitin-conjugating enzymes (UBE2G1 and UBE2G2) to polyubiquitylate and degrade CDT1 post-radiation; this is distinct from the UBCH8 E2 used for p21 and Set8 degradation by the same E3. Proteomic screen for CDT2-associated proteins, siRNA screen, in vitro ubiquitination assay Molecular and cellular biology High 21628527
2011 p97 AAA(+)-ATPase and its cofactor UFD1 are required for proteasome-dependent removal of ubiquitinated CDT1 from chromatin and its subsequent degradation both in vivo and in Xenopus egg extracts; nucleotide excision repair factors promote CDT1 destruction by recruiting PCNA to damaged DNA. Genome-wide siRNA screen, Xenopus egg extract in vitro system, chromatin fractionation Molecular cell High 21981919
2011 CDT1 stabilization by inhibition of NEDD8-activating enzyme (MLN4924) is the critical event triggering DNA rereplication; CDT1 knockdown suppresses MLN4924-induced rereplication, establishing CDT1 as the key cullin substrate mediating this effect. siRNA knockdown of CDT1, cell cycle analysis, MLN4924 treatment Cancer research High 21159650
2011 CDT1 is dynamically recruited to laser-induced DNA damage sites in G1 cells within seconds; recruitment requires PCNA (which is immobile at damage sites); Cdt2 and CUL4/DDB1 also accumulate at damage sites, preceding CDT1 degradation. Laser nanosurgery, live-cell fluorescence microscopy, FRAP, reaction-diffusion modeling Journal of cell science High 21224399
2011 NMR structure of the human Cdt1(410-440)/MCM6(708-821) complex reveals charge complementarity as the key determinant; mutations disrupting this interface abolish MCM2-7 chromatin loading and cause cell death in yeast. NMR structure, mutagenesis, chromatin loading assay, yeast viability assay Nucleic acids research High 22140117
2012 FOXO3 forms a protein complex with CDT1, blocking its interaction with DDB1 and PCNA; FOXO3 depletion accelerates CDT1 proteolysis in unperturbed cells and impairs S-phase entry, establishing FOXO3 as a positive regulator that stabilizes CDT1. Co-immunoprecipitation, siRNA knockdown, cell cycle analysis Proceedings of the National Academy of Sciences of the United States of America Medium 22451935
2014 SCF-FBXO31 E3 ligase interacts with CDT1 and promotes its ubiquitylation and degradation specifically in G2 phase, independently of previously known CUL4 and SCF-Skp2 pathways; FBXO31 depletion stabilizes CDT1 and causes re-replication. Co-immunoprecipitation, ubiquitination assay, cell cycle synchronization, re-replication assay The Journal of biological chemistry Medium 24828503
2015 GRWD1 binds CDT1 and is required for MCM loading; GRWD1 has histone-binding activity, localizes to replication origins in G1 in a CDC6- and Cdt1-dependent manner, and regulates chromatin openness at specific loci. Co-immunoprecipitation, ChIP-seq, FAIRE-seq, siRNA knockdown, MCM loading assay Nucleic acids research Medium 25990725
2016 USP37 deubiquitinase directly interacts with CDT1 and de-ubiquitinates it in vivo; USP37 overexpression stabilizes CDT1 while USP37 knockdown destabilizes it during G1/S; USP37 knockdown reduces MCM chromatin loading and DNA replication fork speed. Co-immunoprecipitation, ubiquitination assay, MCM loading assay, DNA fiber assay, siRNA knockdown Molecular oncology Medium 27296872
2017 Cryo-EM structure of the yeast ORC-Cdc6-Cdt1-Mcm2-7 (OCCM) complex at 3.9 Å shows that Cdt1 adopts a three-domain configuration embracing Mcm2, Mcm4, and Mcm6 (nearly half the hexamer); the Cdt1 C-terminal domain extends to the Mcm6 WHD which contacts Orc4 WHD; Cdt1 stabilizes and orients Mcm2-7 for loading. Cryo-EM structure determination, cross-linking mass spectrometry Nature structural & molecular biology High 28191893
2017 Cryo-EM structures of the yeast MCM hexamer and Cdt1-MCM heptamer show both form left-handed coil structures with a gap between Mcm5 and Mcm2; Cdt1 wraps around the N-terminal regions of Mcm2, Mcm6, and Mcm4 to stabilize the open-ring complex as a precursor to the MCM double hexamer. Cryo-EM structure determination Nature structural & molecular biology High 28191894
2021 SLFN11 promotes CDT1 degradation in response to replicative DNA damage by binding to DDB1 of the CUL4(CDT2) E3 ubiquitin ligase at replication forks; the C-terminus and ATPase domain of SLFN11 are required for DDB1 binding and CDT1 degradation; absence of SLFN11 retards CDT1 degradation, enabling replication reactivation under DNA damage. Co-immunoprecipitation, siRNA knockdown, protein stability assays, domain mutagenesis Proceedings of the National Academy of Sciences of the United States of America High 33536335
2023 CDT1 inhibits CMG helicase activity at replication forks during an overlap period in early S phase when CDT1 is still present after origin firing; CDT1 degradation by CRL4(CDT2) is required for DNA synthesis to commence; this reveals that licensing and firing overlap and cells separate licensing from DNA synthesis rather than strictly separating licensing from firing. Quantitative single-cell microscopy, in vitro reconstituted human DNA replication with purified proteins Molecular cell High 36608667

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2000 Inhibition of eukaryotic DNA replication by geminin binding to Cdt1. Science (New York, N.Y.) 603 11125146
2000 The Cdt1 protein is required to license DNA for replication in fission yeast. Nature 376 10766248
2006 Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cul4, target human Cdt1 for proteolysis. The EMBO journal 327 16482215
1990 Functional studies of nonpeptide angiotensin II receptor subtype-specific ligands: DuP 753 (AII-1) and PD123177 (AII-2). The Journal of pharmacology and experimental therapeutics 323 2243344
2004 Targeted ubiquitination of CDT1 by the DDB1-CUL4A-ROC1 ligase in response to DNA damage. Nature cell biology 314 15448697
1991 Inferring the evolutionary histories of the Adh and Adh-dup loci in Drosophila melanogaster from patterns of polymorphism and divergence. Genetics 265 1673107
2003 Radiation-mediated proteolysis of CDT1 by CUL4-ROC1 and CSN complexes constitutes a new checkpoint. Nature cell biology 256 14578910
2005 PCNA functions as a molecular platform to trigger Cdt1 destruction and prevent re-replication. Nature cell biology 253 16362051
2010 NEDD8-targeting drug MLN4924 elicits DNA rereplication by stabilizing Cdt1 in S phase, triggering checkpoint activation, apoptosis, and senescence in cancer cells. Cancer research 250 21159650
2001 The human licensing factor for DNA replication Cdt1 accumulates in G1 and is destabilized after initiation of S-phase. The Journal of biological chemistry 225 11555648
2002 Interdependent nuclear accumulation of budding yeast Cdt1 and Mcm2-7 during G1 phase. Nature cell biology 219 11836525
2003 The SCF(Skp2) ubiquitin ligase complex interacts with the human replication licensing factor Cdt1 and regulates Cdt1 degradation. The Journal of biological chemistry 208 12840033
2006 PCNA is a cofactor for Cdt1 degradation by CUL4/DDB1-mediated N-terminal ubiquitination. The Journal of biological chemistry 205 16407252
1994 cdt1 is an essential target of the Cdc10/Sct1 transcription factor: requirement for DNA replication and inhibition of mitosis. The EMBO journal 201 8313888
2004 Replication-dependent destruction of Cdt1 limits DNA replication to a single round per cell cycle in Xenopus egg extracts. Genes & development 164 15598982
1990 Nonpeptide angiotensin II receptor antagonists. Studies with EXP9270 and DuP 753. Hypertension (Dallas, Tex. : 1979) 159 2351436
2004 Cyclin-dependent kinases phosphorylate human Cdt1 and induce its degradation. The Journal of biological chemistry 151 15004027
1990 Proximal nephron and renal effects of DuP 753, a nonpeptide angiotensin II receptor antagonist. Kidney international 148 2232490
2006 DTL/CDT2 is essential for both CDT1 regulation and the early G2/M checkpoint. Genes & development 140 17085480
2017 Structural basis of Mcm2-7 replicative helicase loading by ORC-Cdc6 and Cdt1. Nature structural & molecular biology 139 28191893
2011 Inhibition of NEDD8-activating enzyme induces rereplication and apoptosis in human tumor cells consistent with deregulating CDT1 turnover. Cancer research 139 21487042
2008 The inv dup (15) or idic (15) syndrome (Tetrasomy 15q). Orphanet journal of rare diseases 125 19019226
2004 Structural basis for inhibition of the replication licensing factor Cdt1 by geminin. Nature 120 15286659
2004 Human geminin promotes pre-RC formation and DNA replication by stabilizing CDT1 in mitosis. The EMBO journal 118 15257290
2004 Cdt1 downregulation by proteolysis and geminin inhibition prevents DNA re-replication in Xenopus. The EMBO journal 116 15616577
2002 Mouse geminin inhibits not only Cdt1-MCM6 interactions but also a novel intrinsic Cdt1 DNA binding activity. The Journal of biological chemistry 116 12192004
2007 Orc6 is required for dynamic recruitment of Cdt1 during repeated Mcm2-7 loading. Genes & development 115 18006685
2007 Nuclear accumulation of cyclin D1 during S phase inhibits Cul4-dependent Cdt1 proteolysis and triggers p53-dependent DNA rereplication. Genes & development 114 18006686
1993 Uniparental disomy explains the occurrence of the Angelman or Prader-Willi syndrome in patients with an additional small inv dup(15) chromosome. Journal of medical genetics 110 8411071
1991 In vivo pharmacology of DuP 753. American journal of hypertension 107 1854454
2002 Oncogenic potential of the DNA replication licensing protein CDT1. Oncogene 105 11850834
2006 An evolutionarily conserved function of proliferating cell nuclear antigen for Cdt1 degradation by the Cul4-Ddb1 ubiquitin ligase in response to DNA damage. The Journal of biological chemistry 98 16407242
2011 A genome-wide screen identifies p97 as an essential regulator of DNA damage-dependent CDT1 destruction. Molecular cell 97 21981919
2017 Open-ringed structure of the Cdt1-Mcm2-7 complex as a precursor of the MCM double hexamer. Nature structural & molecular biology 94 28191894
2006 Deregulation of Cdt1 induces chromosomal damage without rereplication and leads to chromosomal instability. Journal of cell science 92 16835273
2002 Geminin becomes activated as an inhibitor of Cdt1/RLF-B following nuclear import. Current biology : CB 88 11967157
1990 [3H]DUP 753, a highly potent and specific radioligand for the angiotensin II-1 receptor subtype. Biochemical and biophysical research communications 88 2244903
1991 Nonpeptide angiotensin II receptor antagonists. Studies with DuP 753 and EXP3174 in dogs. European journal of pharmacology 87 1748155
2003 The regulated association of Cdt1 with minichromosome maintenance proteins and Cdc6 in mammalian cells. The Journal of biological chemistry 86 14672932
2005 Degradation of Cdt1 during S phase is Skp2-independent and is required for efficient progression of mammalian cells through S phase. The Journal of biological chemistry 85 15855168
1987 Inhibition of pyrimidine de novo synthesis by DUP-785 (NSC 368390). Investigational new drugs 85 2822596
2004 Regulation of Geminin and Cdt1 expression by E2F transcription factors. Oncogene 84 14990995
2006 DDB1 maintains genome integrity through regulation of Cdt1. Molecular and cellular biology 79 16940174
1991 DuP 753 is a specific antagonist for the angiotensin receptor. Hypertension (Dallas, Tex. : 1979) 79 1672862
2005 The inv dup(15) or idic(15) syndrome: a clinically recognisable neurogenetic disorder. Brain & development 75 16023554
2006 A Cdt1-geminin complex licenses chromatin for DNA replication and prevents rereplication during S phase in Xenopus. The EMBO journal 74 17124498
1996 Cytogenetic and molecular analysis of inv dup(15) chromosomes observed in two patients with autistic disorder and mental retardation. American journal of medical genetics 71 8669450
1991 In vitro pharmacology of DuP 753. American journal of hypertension 70 1854453
2009 Acetylation/deacetylation modulates the stability of DNA replication licensing factor Cdt1. The Journal of biological chemistry 68 19276081
2009 Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing. Proceedings of the National Academy of Sciences of the United States of America 67 19906994
2008 MCM9 binds Cdt1 and is required for the assembly of prereplication complexes. Molecular cell 65 18657502
2004 Rapid degradation of Cdt1 upon UV-induced DNA damage is mediated by SCFSkp2 complex. The Journal of biological chemistry 64 15102855
2021 SLFN11 promotes CDT1 degradation by CUL4 in response to replicative DNA damage, while its absence leads to synthetic lethality with ATR/CHK1 inhibitors. Proceedings of the National Academy of Sciences of the United States of America 62 33536335
2007 Cdt1 associates dynamically with chromatin throughout G1 and recruits Geminin onto chromatin. The EMBO journal 62 17318181
1985 Further delineation of the dup(3q) syndrome. American journal of medical genetics 61 4050847
2007 Cdt1 degradation to prevent DNA re-replication: conserved and non-conserved pathways. Cell division 60 17565698
2007 Identification of novel human Cdt1-binding proteins by a proteomics approach: proteolytic regulation by APC/CCdh1. Molecular biology of the cell 59 18162579
2002 Essential role of human CDT1 in DNA replication and chromatin licensing. Journal of cell science 59 11896191
2000 Characteristics of two cases with dup(15)(q11.2-q12): one of maternal and one of paternal origin. Genetics in medicine : official journal of the American College of Medical Genetics 58 11397326
1991 Effects of the nonpeptide angiotensin II receptor antagonist DuP 753 on blood pressure and renal functions in spontaneously hypertensive PH dogs. American journal of hypertension 58 1854460
2010 Chromatin unfolding by Cdt1 regulates MCM loading via opposing functions of HBO1 and HDAC11-geminin. Cell cycle (Georgetown, Tex.) 57 20980834
2005 Functional domains of the Xenopus replication licensing factor Cdt1. Nucleic acids research 56 15653632
2004 Molecular characterization of inv dup del(8p): analysis of five cases. American journal of medical genetics. Part A 53 15214003
1990 Analysis of DNA polymorphisms suggests that most de novo dup(21q) chromosomes in patients with Down syndrome are isochromosomes and not translocations. American journal of human genetics 53 1978562
2010 Characterization and structure determination of the Cdt1 binding domain of human minichromosome maintenance (Mcm) 6. The Journal of biological chemistry 52 20202939
2008 Cdt1 and Geminin in cancer: markers or triggers of malignant transformation? Frontiers in bioscience : a journal and virtual library 51 18508524
1992 Cardiac angiotensin receptors: effects of selective angiotensin II receptor antagonists, DUP 753 and PD 121981, in rabbit heart. The Journal of pharmacology and experimental therapeutics 51 1602398
2015 Cdt1-binding protein GRWD1 is a novel histone-binding protein that facilitates MCM loading through its influence on chromatin architecture. Nucleic acids research 50 25990725
2014 SCF-FBXO31 E3 ligase targets DNA replication factor Cdt1 for proteolysis in the G2 phase of cell cycle to prevent re-replication. The Journal of biological chemistry 48 24828503
2002 Regulation of CDC6, geminin, and CDT1 in human cells that undergo polyploidization. Molecular biology of the cell 45 12429841
1994 The pharmacokinetics and pharmacodynamics of the angiotensin II receptor antagonist losartan potassium (DuP 753/MK 954) in the dog. The Journal of pharmacology and experimental therapeutics 45 8138932
1991 Pharmacology of DuP 532, a selective and noncompetitive AT1 receptor antagonist. The Journal of pharmacology and experimental therapeutics 45 1941632
2006 Molecular cytogenetic analysis of de novo dup(5)(q35.2q35.3) and review of the literature of pure partial trisomy 5q. American journal of medical genetics. Part A 44 16770806
2011 Selective ubiquitylation of p21 and Cdt1 by UBCH8 and UBE2G ubiquitin-conjugating enzymes via the CRL4Cdt2 ubiquitin ligase complex. Molecular and cellular biology 43 21628527
2004 Geminin has dimerization, Cdt1-binding, and destruction domains that are required for biological activity. The Journal of biological chemistry 42 15308655
2011 JNK1 phosphorylation of Cdt1 inhibits recruitment of HBO1 histone acetylase and blocks replication licensing in response to stress. Molecular cell 41 21856198
2005 Expression of the licensing factors, Cdt1 and Geminin, in human colon cancer. International journal of oncology 40 16273206
2004 Cell cycle regulation of the licensing activity of Cdt1 in Xenopus laevis. Experimental cell research 40 15051497
1992 Chronic captopril and losartan (DuP 753) administration in rats with high-output heart failure. The American journal of physiology 40 1415610
1988 DUP 785 (NSC 368390): schedule-dependency of growth-inhibitory and antipyrimidine effects. Biochemical pharmacology 40 2840910
2021 SIRT3 promotion reduces resistance to cisplatin in lung cancer by modulating the FOXO3/CDT1 axis. Cancer medicine 38 33655712
2016 USP37 deubiquitinates Cdt1 and contributes to regulate DNA replication. Molecular oncology 37 27296872
2011 Structural insights into the Cdt1-mediated MCM2-7 chromatin loading. Nucleic acids research 37 22140117
2008 Cdt1 and Cdc6 are destabilized by rereplication-induced DNA damage. The Journal of biological chemistry 37 18617514
2007 Licensing regulators Geminin and Cdt1 identify progenitor cells of the mouse CNS in a specific phase of the cell cycle. Neuroscience 37 17533120
2011 Dynamic recruitment of licensing factor Cdt1 to sites of DNA damage. Journal of cell science 36 21224399
2005 Licensing for DNA replication requires a strict sequential assembly of Cdc6 and Cdt1 onto chromatin in Xenopus egg extracts. Nucleic acids research 35 15687385
1993 Effect of chronic treatment with losartan potassium (DuP 753) on the elevation of blood pressure during chronic exposure of rats to cold. Pharmacology 33 8483966
2007 Cdt1 and geminin: role during cell cycle progression and DNA damage in higher eukaryotes. Frontiers in bioscience : a journal and virtual library 32 17127409
2012 Regulation of cell cycle progression by forkhead transcription factor FOXO3 through its binding partner DNA replication factor Cdt1. Proceedings of the National Academy of Sciences of the United States of America 31 22451935
1995 A familial Xp+ chromosome, dup (Xq26.3-->qter). Journal of medical genetics 31 8592335
1993 Acute and chronic effects of losartan (DuP 753) on blood pressure and vascular reactivity in normotensive rats. Clinical and experimental hypertension (New York, N.Y. : 1993) 31 8467311
2020 Efficient production of large deletion and gene fragment knock-in mice mediated by genome editing with Cas9-mouse Cdt1 in mouse zygotes. Methods (San Diego, Calif.) 29 32334080
2013 Geminin deploys multiple mechanisms to regulate Cdt1 before cell division thus ensuring the proper execution of DNA replication. Proceedings of the National Academy of Sciences of the United States of America 29 23836640
2008 Cdt1 forms a complex with the minichromosome maintenance protein (MCM) and activates its helicase activity. The Journal of biological chemistry 29 18606811
2000 Isolation and molecular analysis of inv dup(15) and construction of a physical map of a common breakpoint in order to elucidate their mechanism of formation. Chromosoma 26 11151680
2023 CDT1 inhibits CMG helicase in early S phase to separate origin licensing from DNA synthesis. Molecular cell 25 36608667
2013 The link between intraneuronal N-truncated amyloid-β peptide and oxidatively modified lipids in idiopathic autism and dup(15q11.2-q13)/autism. Acta neuropathologica communications 25 24252310
2012 Evidence for a role of Arabidopsis CDT1 proteins in gametophyte development and maintenance of genome integrity. The Plant cell 25 22773747
2021 Regulation of DNA Replication Licensing and Re-Replication by Cdt1. International journal of molecular sciences 23 34068957