| 1997 |
AND-1 (WDHD1) was identified and characterized as a novel nuclear DNA-binding protein containing an N-terminal WD-repeat domain and a C-terminal HMG-box domain. It binds four-way junction DNA, exists as an oligomer (likely homodimer), and localizes to the interchromatinic space during interphase but disperses to the cytoplasm during mitosis. |
cDNA cloning, monoclonal antibody characterization, DNA affinity chromatography, EMSA, immunolocalization |
Journal of cell science |
Medium |
9175701
|
| 1992 |
CTF4/CHL15 (yeast ortholog of WDHD1) encodes a ~105 kDa protein that associates in vitro with DNA polymerase alpha; ctf4 mutants show elevated sister chromatid loss and recombination but not elevated point mutation rates, and the C-terminal helix-loop-helix domain is essential for CTF4 function. |
Yeast genetics, chromosome segregation assays, in vitro DNA polymerase alpha association, sequence analysis, frameshift mutagenesis |
Molecular and cellular biology |
Medium |
1341195
|
| 1999 |
Yeast CTF4 genetically interacts with DNA2 and POL1 (Pol alpha subunit); ctf4 deletion is synthetically lethal with DNA damage-sensitive dna2 alleles, placing Ctf4 in a pathway with Dna2 and Pol alpha for lagging strand synthesis or repair. |
Yeast genetic epistasis, synthetic lethality analysis, allele construction |
Genetics |
Medium |
10101169
|
| 2001 |
Yeast CTF4 is required for sister chromatid cohesion; ctf4 null mutants exhibit cohesion failure and preanaphase accumulation dependent on the spindle assembly checkpoint, placing CTF4 functionally at the replication fork in the cohesion establishment pathway. |
Yeast genetics, sister chromatid cohesion assay, checkpoint epistasis |
Molecular and cellular biology |
High |
11287619
|
| 2002 |
Fission yeast Mcl1 (ortholog of CTF4/WDHD1) is required for chromosome replication, sister chromatid cohesion, and segregation; mcl1-1 is synthetically lethal with checkpoint mutations rad3Δ and rad26Δ, and interacts genetically with replication components. |
Fission yeast genetics, pulsed-field electrophoresis, synthetic lethality, overexpression analysis |
Eukaryotic cell |
Medium |
12455694
|
| 2004 |
Yeast CTF4 (polymerase-alpha-associated protein) is required for efficient sister chromatid cohesion in both mitosis and meiosis; ctf4Δ cells show massive meiosis II non-disjunction and loss of centromeric cohesion despite normal cohesin association with centromeric DNA. |
Yeast genetics, cohesion assays, cytological analysis of meiosis |
Journal of cell science |
Medium |
15226378
|
| 2007 |
Human AND-1/CTF4 (WDHD1) interacts with Mcm10 (which associates with MCM2-7) and with the p180 subunit of DNA polymerase alpha. In Xenopus egg extracts, AND-1 loads on chromatin after Mcm10, concurrently with Pol alpha, and is required for efficient DNA synthesis. Mcm10 is required for chromatin loading of AND-1 and Pol alpha; disrupting the Mcm10-AND-1 interaction inhibits DNA synthesis. |
Co-immunoprecipitation, Xenopus egg extract DNA replication assay, chromatin fractionation, inhibitory antibody, siRNA knockdown in mammalian cells |
Genes & development |
High |
17761813
|
| 2009 |
Yeast Ctf4 couples the MCM2-7 helicase to DNA polymerase alpha within the replisome progression complex (RPC); Ctf4 and GINS together are crucial for this coupling. In ctf4Δ cells, Pol alpha is unstable and barely detectable at replication forks, while helicase (and Pol epsilon) quantities are unchanged but their localizations are abnormally dispersed. Cells lacking both Ctf4 and Mrc1 experience chronic DNA damage checkpoint activation and fail to complete the cell cycle. |
Yeast genetics, ChIP, in vitro interaction assays between GINS and Pol alpha mediated by Ctf4, immunoprecipitation, double mutant analysis |
The EMBO journal |
High |
19661920
|
| 2009 |
Yeast Ctf4 mediates the in vitro interaction between GINS and DNA Pol alpha-primase; in ctf4Δ mutants, the MCM-GINS–Pol alpha interaction is specifically destabilized, Pol alpha becomes unstable at replication forks, and the replication checkpoint is activated. Tof1 and Mrc1 are not required for this interaction. |
Yeast genetics, in vitro protein interaction assay, ChIP, immunoblot |
Genes to cells |
Medium |
19496828
|
| 2009 |
Human AND-1/CTF4 (WDHD1) is required for assembly of the CMG (Cdc45-Mcm2-7-GINS) complex in human cells; along with RecQL4 and Mcm10, AND-1 is needed for stable CMG interactions observed only after G1/S transition and dependent on CDK and Cdc7 kinase activities. |
Bimolecular fluorescence complementation (BiFC) in HeLa cells, siRNA depletion, CDK and Cdc7 kinase inhibition |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
19805216
|
| 2009 |
Human AND-1 (WDHD1) localizes adjacent to replication foci in S phase, is phosphorylated in response to replication arrest in a checkpoint kinase (ATM, ATR, Cdc7)-dependent manner, interacts with cohesin proteins Smc1, Smc3, and Rad21/Scc1, and is required for homologous recombination repair. Depletion of AND-1 increases DNA damage, delays S-phase progression, compromises checkpoint activation, and decreases Chk1 protein levels. |
Immunofluorescence, co-immunoprecipitation, siRNA knockdown, HR repair assay (I-SceI DSB), immunoblot, flow cytometry |
The Journal of biological chemistry |
Medium |
19439411
|
| 2010 |
Purified human CTF4 (WDHD1) exists as a dimer; the SepB domain determines dimeric structure. hCtf4 binds preferentially to DNA template-primer structures, directly interacts with DNA polymerases alpha, delta, and epsilon, and markedly stimulates Pol alpha and Pol epsilon activities in vitro. siRNA depletion of hCtf4 in HeLa cells causes G1/S arrest and slower DNA replication. |
Protein purification, in vitro DNA polymerase stimulation assay, EMSA/DNA binding, co-immunoprecipitation, siRNA knockdown, DNA fiber analysis |
The Journal of biological chemistry |
High |
20089864
|
| 2010 |
Xenopus Dna2 forms a complex with And-1/Ctf4 and Mcm10 at replication forks (demonstrated by co-immunoprecipitation from egg extracts), and is recruited to DNA after replication origin licensing. |
Xenopus egg extract, co-immunoprecipitation, immunofluorescence/co-localization |
Cell cycle |
Medium |
20237432
|
| 2011 |
Human WDHD1 associates with centromeres in a cell cycle-dependent manner (mid-to-late S phase), and its down-regulation compromises HP1alpha localization to pericentric heterochromatin, alters epigenetic marks at pericentric chromatin, disrupts mitosis, and impairs generation of centromeric satellite small non-coding RNAs by reducing Dicer association with centromeric RNA. |
ChIP, immunofluorescence, siRNA knockdown, chromatin fractionation, RNA analysis |
Nucleic acids research |
Medium |
21266480
|
| 2011 |
AND-1 (WDHD1) forms a complex with both histone H3 and histone acetyltransferase Gcn5; AND-1 down-regulation causes Gcn5 protein degradation, reducing H3K9 and H3K56 acetylation. AND-1 overexpression stabilizes Gcn5 through protein-protein interaction. |
Co-immunoprecipitation, siRNA knockdown, immunoblot, overexpression |
Oncogene |
Medium |
21725360
|
| 2012 |
Mimosine prevents chromatin binding of Ctf4/AND-1 in HeLa cells, arresting the cell cycle in G1 before S-phase entry via a Hif-1alpha-dependent increase in p27; depletion of Hif-1alpha restores Ctf4 chromatin binding and allows S-phase entry even in the presence of mimosine. |
Chromatin fractionation, siRNA knockdown, cell cycle analysis, flow cytometry |
Cell cycle |
Medium |
22374673
|
| 2013 |
Human CTF4 (WDHD1) interacts with the CMG (Cdc45-Mcm2-7-GINS) complex; the hCtf4-CMG complex was isolated by in vitro interaction of purified proteins, by co-infection of insect cells, and from HeLa cell chromatin. The complex contains homodimeric hCtf4 and monomeric CMG. The hCtf4-CMG complex retains DNA helicase activity that is more salt-resistant than CMG alone. The stability of hCtf4-CMG depends on interactions between hCtf4 and multiple CMG components. |
Purified protein interaction, Sf9 co-infection/co-purification, HeLa chromatin immunoprecipitation, helicase assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
24255107
|
| 2014 |
Yeast Ctf4 self-associates as a constitutive disk-shaped homotrimer via a C-terminal beta-propeller (SepB) domain fused to a helical extension. Pol alpha catalytic subunit and GINS subunit Sld5 share a conserved Ctf4-interacting peptide (CIP-box) that docks onto the helical extension of a Ctf4 protomer; one Ctf4 trimer can simultaneously bind both Pol alpha and GINS, enabling coupling of two Pol alpha molecules to one CMG helicase. |
X-ray crystallography, electron microscopy, native mass spectrometry, mutagenesis, in vitro binding assays |
Nature |
High |
24805245
|
| 2015 |
Yeast Pol alpha/Primase/Ctf4 mutants that are proficient in bulk DNA replication are defective in recombination-mediated damage bypass by template switching and have reduced sister chromatid cohesion; these DDT defects are caused by altered ssDNA metabolism and abnormal replication fork topology rather than increased sister chromatid distance. |
Yeast genetics, DNA fiber analysis, electron microscopy of replication intermediates, sister chromatid cohesion assay |
Molecular cell |
High |
25661486
|
| 2015 |
RecQL4 is required for the association of Mcm10 and Ctf4 with replication origins in human cells; physical interactions between RecQL4, Mcm10, and Ctf4 and their origin association require both CDK and DDK activities and are blocked by the DNA damage checkpoint. |
ChIP at replication origins, co-immunoprecipitation, siRNA knockdown, kinase inhibition |
Cell cycle |
Medium |
25602958
|
| 2015 |
In response to replication stress, AND-1 is phosphorylated at T826 by ATR; this phosphorylation causes AND-1 to accumulate at damage sites where it promotes the Claspin-Chk1 interaction, stimulating efficient Chk1 activation by ATR. AND-1 directly binds ssDNA and facilitates Claspin association with ssDNA. AND-1 is required for recovery of stalled forks. |
Phosphorylation site mutagenesis, co-immunoprecipitation, in vitro ssDNA binding assay, siRNA knockdown, immunofluorescence, DNA fiber analysis |
The EMBO journal |
High |
26082189
|
| 2015 |
Histone H3K56 acetylation acts through Ctf4 to uncouple CMG helicase from DNA polymerases under replication stress; the N-terminal domain of Ctf4 interacts with Mms22 (adaptor of the Rtt101-Mms1 E3 ubiquitin ligase), and this interaction is promoted by replicative stress. |
Yeast genetics, genetic epistasis, domain deletion analysis, two-hybrid/co-immunoprecipitation |
Genetics |
Medium |
25697176
|
| 2016 |
Ctf4 recruits the Chl1 helicase to the replisome via a conserved CIP-box interaction motif shared with GINS and Pol alpha; EM analysis visualized a reconstituted Chl1-Ctf4-GINS assembly. Ctf4 interaction (but not Chl1 helicase activity) is required for sister chromatid cohesion, while Chl1 helicase activity is required for replication fork progression under nucleotide depletion. Physical interaction between Chl1 and cohesin occurs during S phase. |
Electron microscopy of reconstituted complex, in vitro binding assays, yeast genetics with separation-of-function mutants, co-immunoprecipitation |
Molecular cell |
High |
27397686
|
| 2016 |
Ctf4 acts as a hub connecting multiple CIP-box proteins (in addition to Pol alpha and GINS) to the CMG helicase; crystallographic analysis classifies CIP-boxes into two groups targeting different sites on Ctf4. Ctf4-dependent recruitment of Dna2 and Tof2 via their CIP-boxes is required for rDNA copy-number regulation but not genome-wide DNA synthesis. |
X-ray crystallography, in vitro binding assays, yeast genetics, CIP-box mutagenesis, genome-wide replication analysis |
Molecular cell |
High |
27397685
|
| 2016 |
WDHD1 (AND-1) plays an important role in E7 (HPV-16)-induced G1 checkpoint abrogation and rereplication; WDHD1 protein steady-state level and half-life are increased in E7-expressing cells, and WDHD1 knockdown reduces E7-induced G1 checkpoint abrogation. |
siRNA knockdown, immunoblot (protein stability), cell cycle analysis, rereplication assay |
Journal of virology |
Medium |
27099318
|
| 2017 |
Human AND-1 (WDHD1) maintains the trimeric structure of yeast Ctf4 via its conserved SepB domain, but interacts with DNA Pol alpha/primase primarily through its C-terminal HMG box, which binds the Pol alpha B subunit at the same site targeted by SV40 T-antigen. AND-1 also possesses a DNA-binding activity via its HMG box that may position Pol alpha on the lagging-strand template. |
Biochemical protein interaction assays, domain mutagenesis, competition with SV40 T-antigen peptide, DNA binding assay |
Open biology |
High |
29167311
|
| 2017 |
Human AND-1 (WDHD1) exists as a homotrimer mediated by the SepB domain; the positively charged groove within SepB provides binding sites for Pol alpha. AND-1 binds DNA in vitro via its C-terminal HMG domain. Unlike yeast Ctf4, human AND-1's recruitment to the CMG complex appears to be mediated by unknown participants other than GINS. |
X-ray crystallography of WD40 and SepB domains, biochemical assays, mutagenesis, DNA binding assay |
The Journal of biological chemistry |
High |
28381552
|
| 2017 |
AND-1/Ctf4 promotes DNA end resection and homologous recombination repair by interacting with CtIP and regulating its recruitment to DNA damage sites. AND-1 localizes to damage sites via the MDC1-RNF8 pathway and is required for sustained ATR-CHK1 signaling and maintenance of intra-S and G2-phase checkpoints. |
Co-immunoprecipitation, siRNA knockdown, immunofluorescence at damage foci, HR repair assay, checkpoint assays |
Nucleic acids research |
Medium |
27940552
|
| 2017 |
AND-1 (WDHD1) forms complexes with CtIP and other repair proteins; AND-1 is recruited to DSB sites dependent on MDC1, BRCA1, and ATM. AND-1 knockdown impairs DSB end resection by reducing CtIP recruitment, reduces Chk1 activation, establishing an MDC1→AND-1→CtIP axis for CtIP-mediated end resection. |
Co-immunoprecipitation, siRNA knockdown, immunofluorescence at damage foci, end resection assay, Chk1 activation assay |
Nucleic acids research |
Medium |
27940557
|
| 2017 |
Yeast Ctf4 prevents genome rearrangements by suppressing DSB formation and end resection at arrested replication forks; in ctf4Δ cells, DSBs form more frequently at arrested forks, undergo end resection, and are repaired by HR prone to rDNA hyper-amplification. |
Yeast genetics, Southern blotting/rDNA copy number, DSB quantification, end resection assay |
Molecular cell |
Medium |
28525744
|
| 2017 |
A stapled peptide encoding the Ctf4-interacting peptide (CIP) of Sld5 binds Ctf4 with submicromolar affinity; crystal structure confirmed the mode of interaction; the stapled peptide displaces Pol alpha from the replisome in yeast extracts. |
Stapled peptide design, crystal structure, fluorescence polarization binding assay, yeast extract replisome disruption assay |
Angewandte Chemie |
High |
28815832
|
| 2017 |
ATR-Chk1 inhibitor-induced unscheduled origin firing in human cells is mediated by Cdc7 kinase through phosphorylations on GINS that induce an association between GINS and AND-1 (WDHD1). |
Proteomics (phosphoproteomics), co-immunoprecipitation, kinase inhibition, siRNA knockdown |
Nature communications |
Medium |
29123096
|
| 2018 |
The Mcm2-Ctf4-Pol alpha axis facilitates transfer of parental histone (H3-H4)2 tetramers to lagging-strand DNA at replication forks; mutations disrupting the Mcm2 histone-binding domain, or Ctf4-Pol alpha connection, result in enrichment of parental histones on leading strand DNA. |
SCAR-seq (strand-specific parental histone enrichment), yeast genetics with separation-of-function mutations, ChIP |
Molecular cell |
High |
30244834
|
| 2018 |
AND-1 (WDHD1) protects replication forks from resection; depletion of AND-1 using an inducible degron causes fork speed slow-down, accumulation of long ssDNA gaps at the fork junction, and conversion of these to DSBs in G2 via MRE11 nuclease activity. The WD40 domain of AND-1 is specifically required for fork protection and proliferation, while the HMG box is required for fast replication but not proliferation. |
Auxin-inducible degron (AND-1 depletion), DNA fiber analysis, electron microscopy of replication intermediates, MRE11 inhibitor (mirin), domain-specific mutants |
Nature communications |
High |
30082684
|
| 2019 |
Ctf4 tightly dimerizes CMG helicase via an interface involving Psf2, Cdc45, and Sld5; Ctf4 binds only one Pol alpha-primase per trimer, suggesting a replication factory model where two CMG helicases and one Pol alpha-primase are organized by one Ctf4 trimer. Cryo-EM reveals two CMGs nearly facing each other with lagging strands toward the center. |
Cryo-EM structure, in vitro reconstitution, pulldown assays |
eLife |
High |
31589141
|
| 2019 |
Human CST (CTC1-STN1-TEN1) promotes AND-1/Ctf4 chromatin association and replisome assembly; CST directly interacts with AND-1 and pol alpha, enhancing their chromatin association independently of exogenous replication stress. |
Co-immunoprecipitation, chromatin fractionation, siRNA knockdown, origin licensing assays |
Life science alliance |
Medium |
30979824
|
| 2020 |
CryoEM structure of human CMG bound to AND-1 (CMGA) reveals the AND-1 trimer docks onto the helicase assembly formed by Cdc45 and GINS via one beta-propeller domain of its trimerization region; in the CMGA architecture, AND-1 is closely positioned to fork DNA while its CIP-binding helical domains remain available for partner protein recruitment. |
CryoEM structure at 3.3 Å resolution |
Nucleic acids research |
High |
32453425
|
| 2020 |
Yeast Sen1 (transcription termination helicase) associates with the replisome via Ctf4 and Mrc1; the N-terminus of Sen1 binds Ctf4 and Mrc1. A separation-of-function mutant (sen1-3) that abolishes replisome binding without affecting transcription termination shows increased genome instability and recombination. |
Yeast two-hybrid, co-immunoprecipitation, separation-of-function mutagenesis, genetics |
Cell reports |
Medium |
32075754
|
| 2020 |
Ctf4-mediated Pol alpha recruitment for lagging-strand initiation and for origin firing are both sensitive to Pol alpha abundance, but lagging-strand initiation is specifically more dependent on Ctf4-Pol alpha interaction when Pol alpha is limiting, suggesting Pol alpha associates distributively at the fork. |
Yeast genetics with titratable Pol alpha levels, Okazaki fragment analysis, origin firing efficiency measurement, Ctf4-Pol alpha interaction mutants |
PLoS genetics |
Medium |
32379761
|
| 2020 |
WDHD1 (AND-1) acts as an E3 ubiquitin ligase that promotes ubiquitination of MAPRE2 in the nucleus; WDHD1 and MAPRE2 interact by co-immunoprecipitation, and WDHD1 overexpression leads to MAPRE2 degradation conferring cisplatin resistance in lung adenocarcinoma cells. |
Co-immunoprecipitation, ubiquitination assay, WDHD1 knockout, protein stability assay, xenograft |
Frontiers in oncology |
Medium |
32426268
|
| 2021 |
STAT3 transcription factor binds the promoter/upstream regulatory region of WDHD1 gene and activates its transcription; WDHD1 mediates the DNA replication function of STAT3 (STAT3 knockdown reduces DNA replication, which is rescued by WDHD1 expression). |
ChIP (STAT3 at WDHD1 promoter), siRNA knockdown, ectopic expression, DNA replication assay |
Cell & bioscience |
Medium |
33413624
|
| 2021 |
Two AND-1 inhibitors (bazedoxifene acetate and compound CH3) promote AND-1 degradation by disrupting AND-1 polymerization through direct interaction with the WD40 domain; depolymerization promotes interaction with E3 ligase CUL4B, leading to AND-1 ubiquitination and degradation. |
High-throughput screen, in vitro binding assay (WD40 domain interaction), co-immunoprecipitation (CUL4B interaction), ubiquitination assay, in vitro and in vivo cancer growth assays |
Clinical and translational medicine |
Medium |
34923765
|
| 2022 |
AND-1 (WDHD1) is phosphorylated at T826 by ATR in response to interstrand crosslinks (ICLs); this phosphorylation triggers an intramolecular change that promotes AND-1 interaction with FANCM/FAAP24 complex, recruiting it to ICL-stalled forks to activate Fanconi anemia signaling. |
Co-immunoprecipitation, phosphorylation site mutagenesis (T826A), ATR inhibition, immunofluorescence at ICL foci, proximity ligation assay |
Cancer research |
High |
35867033
|
| 2024 |
ZBTB16 transcription factor inhibits WDHD1 transcription; ectopic WDHD1 expression reverses ZBTB16-mediated inhibition of lung adenocarcinoma cell proliferation and S-phase cell cycle arrest, placing WDHD1 downstream of ZBTB16 in a regulatory axis. |
Ectopic expression, siRNA/overexpression rescue, cell cycle analysis, xenograft, bioinformatics/promoter analysis |
Oncogene |
Medium |
38654107
|