Affinage

Showing RAD17 — RAD24 is a alias.

RAD17

Cell cycle checkpoint protein RAD17 · UniProt O75943

Length
681 aa
Mass
77.1 kDa
Annotated
2026-06-10
100 papers in source corpus 36 papers cited in narrative 34 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RAD17 is the large subunit of an RFC-like (clamp loader) complex that initiates the ATR-dependent DNA damage and replication checkpoint by loading the PCNA-like 9-1-1 (RAD9-RAD1-HUS1) clamp onto damaged DNA (PMID:11799063, PMID:11907025, PMID:12167163). RAD17 assembles with the four small RFC subunits RFC2-RFC5 into a heteropentameric, ATPase-active complex structurally analogous to RFC, and this assembly and RAD17 chromatin association require its conserved nucleotide-binding motif (PMID:11907025, PMID:12167163, PMID:10660302, PMID:11313455). RPA-coated single-stranded DNA stimulates RAD17-RFC binding to primed and gapped substrates, and unlike replicative RFC the RAD17 complex recognizes recessed 5' dsDNA-ssDNA junctions to load and correctly orient 9-1-1 in an ATP-hydrolysis-dependent manner (PMID:14605214, PMID:12604797, PMID:35819203). Engagement of 9-1-1 is mediated through a RAD1-binding motif in the RAD17 N-terminus, a conserved KYxxL motif in its AAA+ domain, and a polyanionic C-terminal iVERGE element whose phosphorylation by CK2 (Ser667) and CK1δ/ε (Thr670) promotes the RAD17-9-1-1 interaction (PMID:27387238, PMID:28666868, PMID:29902452, PMID:31353086, PMID:36841485). Once loaded, RAD17 and 9-1-1 enable ATR to phosphorylate RAD17 at Ser635/Ser645, a modification required for the G1/S and G2 checkpoints, for recruitment of Claspin, and for sustained Chk1 activation, and RAD17 mediates 9-1-1 association with the ATR activator TopBP1 (PMID:11418864, PMID:11687627, PMID:16885023, PMID:20110345). Beyond ATR signaling, RAD17 supports an ATM branch: ATM phosphorylates RAD17 at Thr622 to drive early MRN complex recruitment to double-strand breaks and homologous recombination repair, and RAD17 is essential for genomic stability, HR, chromosomal integrity, and embryonic viability (PMID:24534091, PMID:12672690, PMID:15297881, PMID:19168994). Checkpoint signaling is reversed when Cdh1/APC ubiquitinates RAD17 for proteasomal degradation, permitting checkpoint termination and cell cycle re-entry, while the deubiquitylase USP20 opposes this turnover to stabilize RAD17 (PMID:20424596, PMID:24923443).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 1999 High

    Established that the checkpoint Rad17/Mec3/Ddc1 proteins form a stable trimeric clamp acting downstream of the separate Rad24 clamp-loader, defining a two-module architecture for checkpoint sensing.

    Evidence Yeast two-hybrid, Co-IP, gel filtration co-sedimentation and dosage suppression in S. cerevisiae

    PMID:9891048

    Open questions at the time
    • Did not define the biochemical loading mechanism
    • Human orthology not yet established
  2. 2000 High

    Showed the checkpoint clamp loader is an RFC variant in which Rad24 (RAD17 ortholog) replaces Rfc1 while retaining the four small RFC subunits, distinguishing it from replicative RFC.

    Evidence Purification to homogeneity, mass spectrometry and reciprocal Co-IP in S. cerevisiae; ATP-binding motif mutagenesis (K115E/K115R) and dosage suppression of rfc5-1

    PMID:10660302 PMID:10913172 PMID:9710632

    Open questions at the time
    • Did not show direct clamp loading onto DNA
    • Human complex composition not confirmed
  3. 2001 High

    Identified RAD17 as a direct ATR/ATM substrate at Ser635/Ser645 whose phosphorylation is required for DNA-damage checkpoints and for damage-inducible association with 9-1-1, linking RAD17 modification to checkpoint output.

    Evidence In vitro ATR/ATM kinase assays, S635A/S645A mutagenesis, Co-IP and G1/S and G2 checkpoint assays in human cells; ATP-binding/chromatin coupling in S. pombe (K118E)

    PMID:11313455 PMID:11418864 PMID:11687627

    Open questions at the time
    • Did not resolve how phosphorylation alters 9-1-1 engagement structurally
    • Order of loading versus phosphorylation unclear
  4. 2002 High

    Demonstrated that RAD17 recruits 9-1-1 onto chromatin after damage and that RAD17-loaded 9-1-1 is required for ATR to recognize its substrates, placing RAD17 upstream as a damage sensor.

    Evidence Chromatin fractionation, Co-IP, siRNA and epistasis in human cells

    PMID:11799063

    Open questions at the time
    • Mechanism of ATR substrate licensing by 9-1-1 not defined
    • DNA structure recognized not yet identified
  5. 2003 High

    Reconstituted the core reaction, showing RPA stimulates RAD17-RFC binding to ssDNA/gapped substrates and that the complex loads 9-1-1 onto partial duplexes via ATP, using 5' junctions unlike replicative RFC.

    Evidence In vitro reconstitution with purified human and yeast proteins, DNA-binding and ATP-hydrolysis/sliding assays

    PMID:12604797 PMID:14605214

    Open questions at the time
    • Atomic geometry of junction recognition not yet resolved
    • In vivo substrate at forks not directly visualized
  6. 2003 High

    Linked RAD17 to active replication, showing constitutive chromatin association and phosphorylation-dependent localization to replication sites and interaction with DNA polymerase epsilon, and that RAD17 loss causes endoreduplication and chromosomal instability.

    Evidence Chromatin fractionation, Co-IP with Pol epsilon, BrdU labeling; conditional human somatic knockout with cytogenetics and ploidy analysis

    PMID:12672690 PMID:14500819

    Open questions at the time
    • Direct functional role of Pol epsilon interaction not defined
    • Mechanism linking checkpoint loss to endoreduplication unresolved
  7. 2004 High

    Established RAD17 ATPase activity as required for chromatin loading and S-phase checkpoint, identified MCM7 as a partner needed for ATR focus formation, and demonstrated RAD17 is essential for embryonic viability and homologous recombination.

    Evidence ATPase-mutant analysis and S-phase checkpoint assays; MCM7 Co-IP and siRNA; targeted mouse/ES cell knockout with HR (gene targeting) and survival assays

    PMID:15235112 PMID:15297881 PMID:15538388

    Open questions at the time
    • How MCM7 stabilizes ATR not mechanistically defined
    • Whether HR role is separable from checkpoint signaling unresolved here
  8. 2008 Medium

    Showed RAD17 and ATR jointly immobilize RAD9 into stable damage foci, providing direct dynamic evidence that RAD17 retains the 9-1-1 clamp at lesions.

    Evidence FRAP, immunofluorescence, ATR siRNA and RAD17(AA) mutant in human cells

    PMID:19020305

    Open questions at the time
    • Single lab
    • Does not define molecular tether retaining 9-1-1
  9. 2010 High

    Defined RAD17 ATP binding/hydrolysis as the switch coupling 9-1-1 loading to TopBP1 chromatin accumulation, and established Cdh1/APC-mediated RAD17 degradation as the mechanism terminating the checkpoint to allow cell cycle re-entry.

    Evidence Xenopus extract ATPase-mutant assays and TopBP1 chromatin binding; ubiquitination assays, Cdh1/APC Co-IP and degradation-resistant mutant in human cells

    PMID:20110345 PMID:20424596

    Open questions at the time
    • Indirect link between 9-1-1 and TopBP1 not fully resolved
    • Signal triggering Cdh1/APC targeting of RAD17 unknown
  10. 2014 High

    Revealed an ATM-branch role for RAD17, with ATM phosphorylation at Thr622 driving MRN recruitment to DSBs and HR, and showed RAD17 supports HR independent of replication checkpoint signaling.

    Evidence Phospho-mapping (MS), T622A mutagenesis, NBS1 Co-IP and HR assays in human cells; gene targeting and sister chromatid exchange in DT40 cells; USP20 stabilization of RAD17

    PMID:19168994 PMID:24534091 PMID:24923443

    Open questions at the time
    • How a clamp loader directly promotes HR strand exchange unresolved
    • Crosstalk between ATM and ATR RAD17 phospho-sites not integrated
  11. 2013 High

    Distinguished RAD17-dependent rapid Chk1 phosphorylation from an Nbs1-dependent ATR mode acting on RPA32, showing RAD17 governs a specific branch of ATR activation at replication-associated breaks.

    Evidence siRNA, in vitro ATR kinase assays and phospho-specific antibodies in Xenopus extracts

    PMID:23684611

    Open questions at the time
    • Branch selection determinants not defined
    • Relative contribution in vivo unquantified
  12. 2019 Medium

    Mapped the molecular determinants of RAD17-9-1-1 contact, defining KYxxL, the C-terminal iVERGE motif, and its constitutive phosphorylation by CK2 (Ser667) and CK1δ/ε (Thr670) as regulators of clamp engagement and ATR-Chk1 signaling.

    Evidence Motif/deletion and phosphomimetic mutagenesis, in vitro CK2 and CK1δ/ε kinase assays, CK2 inhibitor (CX-4945), Co-IP in human cells

    PMID:27387238 PMID:28666868 PMID:29902452 PMID:31353086

    Open questions at the time
    • Single lab series
    • In vivo checkpoint requirement of each site not fully tested
  13. 2023 High

    Provided atomic-resolution mechanism for clamp specificity and orientation, showing how RAD17-RFC recognizes recessed 5' DNA ends and how the RAD17 N-terminus docks onto the RAD1 subunit in competition with RHINO.

    Evidence Cryo-EM of human RAD17-RFC:9-1-1:DNA and 2.1 Å crystal structure of 9-1-1 bound to a RAD17 N-terminal peptide with RHINO competition assays

    PMID:35819203 PMID:36841485

    Open questions at the time
    • Functional consequence of RHINO competition in cells not established
    • Dynamics of loading/unloading cycle not captured

Open questions

Synthesis pass · forward-looking unresolved questions
  • How RAD17 mechanistically promotes homologous recombination beyond loading the 9-1-1 checkpoint clamp, and how the ATM-Thr622 and ATR-Ser635/645 phospho-branches are coordinated, remain unresolved.
  • No reconstitution of a direct RAD17 role in strand exchange
  • Integration of ATM and ATR RAD17 modifications uncharacterized
  • Physiological role of Pol epsilon and MCM7 interactions at unperturbed forks unclear

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140657 ATP-dependent activity 5 GO:0060090 molecular adaptor activity 4 GO:0003677 DNA binding 3 GO:0016787 hydrolase activity 3
Localization
GO:0000228 nuclear chromosome 3 GO:0005634 nucleus 2
Pathway
R-HSA-1640170 Cell Cycle 4 R-HSA-8953897 Cellular responses to stimuli 4 R-HSA-73894 DNA Repair 3 R-HSA-69306 DNA Replication 2
Partners
ATRCLASPINHUS1MCM7NBS1RAD1RAD9TOPBP1
Complex memberships
9-1-1 (RAD9-RAD1-HUS1) clamp (loaded substrate)RAD17-RFC (RAD17-RFC2-5) clamp loader

Evidence

Reading pass · 34 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2002 Human Rad17 recruits the Rad9-Rad1-Hus1 (9-1-1) complex onto chromatin after DNA damage. Rad17 binds chromatin prior to damage and is phosphorylated by ATR on chromatin after damage. The phosphorylation of Rad17 by ATR requires Hus1, suggesting that the 9-1-1 complex recruited by Rad17 enables ATR to recognize its substrates. Rad17 and ATR localize to DNA damage largely independently. Chromatin fractionation, co-immunoprecipitation, siRNA knockdown, epistasis analysis in human cells Genes & development High 11799063
2003 Replication protein A (RPA) stimulates binding of the Rad17-Rfc2-5 complex to single-stranded DNA (ssDNA), primed ssDNA, and gapped DNA structures. RPA facilitates recruitment of the Rad9-Rad1-Hus1 complex by Rad17-Rfc2-5 to primed and gapped DNA in vitro. Unlike RFC, which uses the 3' primer/template junction to load PCNA, Rad17-Rfc2-5 can use both 5' and 3' junctions and prefers gapped DNA structures. In vitro biochemical reconstitution with purified proteins, DNA binding assays, pulldown assays Proceedings of the National Academy of Sciences of the United States of America High 14605214
2001 ATM and ATR phosphorylate human Rad17 at Ser635 and Ser645 in response to genotoxic agents. A phosphorylation-deficient Rad17 mutant (Rad17AA, S635A/S645A) abrogated the DNA-damage-induced G2 checkpoint and sensitized cells to genotoxic stress. The Rad17AA mutant showed no ionizing-radiation-inducible association with hRad1, indicating phosphorylation is required for interaction with the 9-1-1 complex. In vitro kinase assay (ATR/ATM), phosphorylation-site mutagenesis, co-immunoprecipitation, cell cycle analysis by flow cytometry, overexpression in human fibroblasts Nature High 11418864
2001 ATR (but not ATM) phosphorylates human Rad17 at Ser635 and Ser645 in vitro. These sites are phosphorylated in a cell-cycle-regulated manner in undamaged cells (late G1, S, and G2/M, but not early-mid G1). Expression of a Rad17 S635A/S645A double mutant abolished IR-induced G1/S checkpoint activation in MCF-7 cells. In vitro kinase assay with recombinant ATR, synchronized cell cycle analysis, mutagenesis, G1/S checkpoint assay in MCF-7 cells Proceedings of the National Academy of Sciences of the United States of America High 11687627
2003 Phosphorylated Rad17 interacts with Claspin and regulates its phosphorylation. A phosphomutant Rad17AA fails to sustain Chk1 phosphorylation after hydroxyurea withdrawal and fails to recruit Claspin, defining an ATR–Rad17–Claspin–Chk1 signaling cascade for replication stress responses. Co-immunoprecipitation of phospho-Rad17 with Claspin, siRNA knockdown, Chk1 phosphorylation assays, cell cycle analysis in human cells Molecular cell High 16885023
2003 The yeast Rad24-RFC complex (clamp loader) loads the Rad17-Mec3-Ddc1 (9-1-1) clamp onto partial-duplex DNA in an ATP-dependent process in vitro. Upon ATP hydrolysis, the 9-1-1 clamp is released from the clamp loader and can slide across >1 kb of duplex DNA. The 9-1-1 clamp showed no detectable exonuclease activity. In vitro reconstitution with purified yeast proteins, ATP hydrolysis assays, DNA mobility shift assays Proceedings of the National Academy of Sciences of the United States of America High 12604797
2002 Human Rad17 forms a heteropentameric complex with four small RFC subunits (hRad17-RFC) with a cleft structure similar to RFC. The 9-1-1 complex forms a trimeric ring structure similar to PCNA. Rad17-RFC exhibits DNA binding and ATPase activity and binds Rad9-1-1. Purification of recombinant complexes from insect cells, electron microscopy (glycerol spray/low-voltage and platinum shadowing), ATPase assay, pulldown The Journal of biological chemistry / Genes to cells High 11907025 12167163
2022 CryoEM structure of human RAD17-RFC clamp loader bound to human 9-1-1 at a dsDNA-ssDNA junction at a recessed 5'-end was determined. The structure reveals how RAD17 confers specificity for 9-1-1 over PCNA and how the clamp loader specifically recognizes the recessed 5' DNA end, fixing the orientation of 9-1-1 on ssDNA. Cryogenic electron microscopy (cryo-EM) structural determination of human RAD17-RFC:9-1-1:DNA complex Nucleic acids research High 35819203
2004 Human MCM7 was identified as a novel Rad17-interacting protein. Depletion of either Rad17 or MCM7 by siRNA suppressed UV- or aphidicolin-induced Chk1 phosphorylation and abolished UV-induced S-phase checkpoint activation. MCM7-depleted cells were defective for ATR nuclear focus formation after UV, suggesting MCM7 is required for stable recruitment of ATR to damaged DNA. Co-immunoprecipitation, siRNA knockdown, Chk1 phosphorylation assay, immunofluorescence for ATR foci The EMBO journal Medium 15538388
2013 Rad17 is required for the rapid phosphorylation of Chk1 but not for a Nbs1-dependent mode of ATR activation that phosphorylates RPA32 Ser33. These represent two distinct modes of ATR activation at replication-associated DSBs. siRNA knockdown, in vitro ATR kinase assay, phospho-specific antibodies, Xenopus egg extracts Cell reports High 23684611
2014 Rad17 is required for early, MDC1-independent recruitment of the MRN (MRE11-RAD50-NBS1) complex to DSB sites and contributes to ATM activation. ATM phosphorylates Rad17 at a novel Thr622 site, which enables direct interaction of Rad17 with NBS1 and facilitates recruitment of MRN/ATM to DSBs, creating a positive feedback loop. Thr622 phosphorylation is important for MRN/ATM signaling and homologous recombination repair. Co-immunoprecipitation, phospho-site mapping by mass spectrometry, point mutagenesis (T622A), HR repair assay (gene targeting), chromatin fractionation in human cells The EMBO journal High 24534091
2010 Rad17 mediates the interaction of the 9-1-1 complex with the ATR-activating protein TopBP1 in Xenopus egg extracts. ATP binding by Rad17 is essential for 9-1-1 and TopBP1 association. ATP hydrolysis by Rad17 is necessary for loading 9-1-1 onto DNA and the checkpoint-dependent accumulation of TopBP1 on chromatin. A 9-1-1 mutant unable to bind TopBP1 still supports normal TopBP1 chromatin accumulation, suggesting TopBP1 accumulation is Rad17- and 9-1-1-dependent but not through direct 9-1-1-TopBP1 docking. Xenopus egg extract checkpoint assays, Rad17 ATPase mutant analysis, TopBP1 chromatin binding assays, co-immunoprecipitation Molecular biology of the cell High 20110345
2010 Rad17 is ubiquitinated and degraded by the Cdh1/APC ubiquitin ligase after UV radiation in human cells. A degradation-resistant Rad17 mutant prevents termination of checkpoint signaling and blocks re-entry into the cell cycle, demonstrating that Rad17 proteolysis by Cdh1/APC is required for checkpoint termination and recovery from genotoxic stress. Ubiquitination assays, Cdh1/APC co-immunoprecipitation, degradation-resistant mutant overexpression, cell cycle re-entry assays, siRNA knockdown in human primary cells The EMBO journal High 20424596
1999 In S. cerevisiae, Rad17 and Mec3 interact physically in vivo (yeast two-hybrid and co-immunoprecipitation). Ddc1 co-sediments and co-immunoprecipitates with both Rad17 and Mec3, forming a trimeric complex. Rad24 does not associate with Rad17, Mec3, or Ddc1. DDC1 overexpression partially suppresses rad24 deletion phenotypes, placing the Rad17-Mec3-Ddc1 complex downstream of Rad24. Yeast two-hybrid screen, co-immunoprecipitation, gel filtration co-sedimentation, genetic epistasis (dosage suppression) Molecular and cellular biology High 9891048
2000 S. cerevisiae Rad24 forms an RFC-like complex with the four small RFC subunits Rfc2, Rfc3, Rfc4, and Rfc5 (but not Rfc1), identified by purification and mass spectrometry. Reciprocal co-immunoprecipitation confirmed these interactions. Biochemical purification to homogeneity, mass spectrometry, reciprocal co-immunoprecipitation Current biology High 10660302
1998 S. cerevisiae Rad24 physically interacts with RFC subunits Rfc2 and Rfc5 and co-sediments with Rfc5. The conserved NTP-binding motif (Lys115) of Rad24 is required for its interaction with RFC proteins and for checkpoint function. RAD24 overexpression suppresses the checkpoint defect of rfc5-1 mutants. Co-immunoprecipitation, co-sedimentation, site-directed mutagenesis (K115E, K115R), dosage suppression, Rad53 phosphorylation assay Molecular and cellular biology High 10913172 9710632
2003 Human Rad17 is chromatin-associated throughout the cell cycle independently of DNA damage. Phosphorylated Rad17 preferentially associates with sites of ongoing DNA replication and interacts with DNA polymerase epsilon (co-immunoprecipitation). Chromatin fractionation, co-immunoprecipitation with DNA polymerase epsilon, BrdU incorporation to mark replication sites, immunofluorescence Nucleic acids research Medium 14500819
2003 RAD17 deletion in human somatic cells leads to acute chromosomal aberrations and endoreduplication at high rates, demonstrating that Rad17 is essential for cell viability and chromosomal stability, linking checkpoint function to ploidy control. Conditional RAD17 knockout in human somatic cells, flow cytometry for ploidy, cytogenetic analysis for chromosomal aberrations Genes & development High 12672690
2014 The deubiquitylase USP20 interacts with Rad17, stabilizes Rad17 protein in steady-state and after DNA damage in a proteasome-dependent manner, and is required for proper Chk1 phosphorylation by ATR. USP20 depletion impairs homologous recombination repair of collapsed replication forks. Co-immunoprecipitation, siRNA knockdown, ubiquitination assays, Chk1 phosphorylation assay, HR repair assay The Journal of biological chemistry Medium 24923443
2001 Fission yeast Rad17 associates with chromatin in response to DNA damage (MMS or ionizing radiation) but dissociates from chromatin during S-phase stall (hydroxyurea). Rad17 complexes in vivo with the Rfc small subunit Rfc2 but not Rfc1. A checkpoint-defective mutant Rad17(K118E) cannot bind ATP, shows reduced chromatin binding, and reduced complex formation with Rfc2, indicating ATP binding is required for chromatin association and RFC complex formation. Chromatin fractionation, co-immunoprecipitation, site-directed mutagenesis (K118E), ATP binding assays in S. pombe Molecular and cellular biology High 11313455
2004 Mouse Rad17 deletion leads to embryonic lethality during early/mid-gestation. Truncated mRad17-expressing ES cells are hypersensitive to DNA-damaging agents and display impaired homologous recombination (strongly reduced gene targeting efficiency), demonstrating a role for Rad17 in DNA damage-dependent recombination beyond checkpoint signaling. Targeted gene deletion in mice and ES cells, clonogenic survival assays with DNA-damaging agents, gene targeting efficiency assay for HR, cell cycle checkpoint assay The EMBO journal High 15297881
2004 HIV-1 Vpr-mediated G2 arrest requires Rad17 and Hus1. Vpr activates ATR signaling, and knockdown/mutation of Rad17 or Hus1 prevents Vpr-induced G2 arrest, phosphorylation of histone H2AX, and formation of γH2AX/BRCA1 nuclear foci. siRNA knockdown of Rad17 and Hus1, flow cytometry for G2 arrest, immunofluorescence for γH2AX and BRCA1 foci in human cells Molecular and cellular biology Medium 15485898
2008 ATR and Rad17 collaborate to regulate Rad9 localization at DNA damage sites. DNA damage or replication stress causes Rad17-dependent immobilization of Rad9 into nuclear foci. Expression of phosphorylation-deficient Rad17(AA) or ATR downregulation reduces the number of cells with Rad9 foci and increases the dynamic behavior of Rad9 within foci (measured by FRAP). Immunofluorescence, FRAP (fluorescence recovery after photobleaching), siRNA knockdown of ATR, overexpression of Rad17(AA) mutant in human cells Journal of cell science Medium 19020305
2004 ATPase activity of Rad17 is required for its chromatin association and for ATR-mediated S-phase checkpoint activation in response to low-dose UV radiation in human cells. ATPase-deficient Rad17 mutant expression, chromatin fractionation, S-phase checkpoint assay (DNA synthesis inhibition) in human cells Molecular cancer research Medium 15235112
2016 A conserved KYxxL motif in the AAA+ domain of Rad17 is essential for interaction with the 9-1-1 complex. Rad17 KYxxL mutants show increased UV-induced Rad17 phosphorylation, suggesting the 9-1-1 complex negatively regulates Rad17 phosphorylation. Nucleotide-binding activity of Rad17 is required for its nuclear localization. Site-directed mutagenesis of KYxxL motif, co-immunoprecipitation, phosphorylation assays by western blot, nuclear localization analysis Biochemical and biophysical research communications Medium 27387238
2017 The polyanionic C-terminal tail of human Rad17 contains a conserved sequence motif (iVERGE) that is required for interaction with the 9-1-1 complex and regulation of ATR-Chk1 signaling. Deletion mutagenesis of C-terminal tail, co-immunoprecipitation with 9-1-1, Chk1 phosphorylation assay Biochemical and biophysical research communications Medium 28666868
2018 Casein kinase 2 (CK2) constitutively phosphorylates Rad17 at Ser667 within the C-terminal iVERGE motif, and this phosphorylation is required for interaction between Rad17 and the 9-1-1 complex. CK2 inhibition by CX-4945/Silmitasertib impairs Rad17-9-1-1 interaction in a Ser667-dependent manner. Site-directed mutagenesis (S667A), CK2 inhibitor treatment, co-immunoprecipitation, in vitro CK2 kinase assay Biochemical and biophysical research communications Medium 29902452
2019 CK1δ/ε phosphorylates Rad17 at Thr670. T670 phosphorylation promotes Ser667 phosphorylation (and vice versa), and phosphomimetic T670 mutations enhance interaction with the 9-1-1 complex, indicating multi-site phosphorylation at the C-terminal iVERGE motif regulates ATR-Chk1 signaling. In vitro CK1δ/ε kinase assay, phosphomimetic mutagenesis, co-immunoprecipitation with 9-1-1 Biochemical and biophysical research communications Medium 31353086
2023 Crystal structure (2.1 Å) of human 9-1-1 bound to a RAD17 N-terminal peptide containing a RAD1-binding motif was determined. The N-terminal region of RAD17 binds specifically to the RAD1 subunit of 9-1-1 via defined interactions. The RAD1-binding motif of RHINO competes with the RAD17 N-terminal region for RAD1 binding, implying functional roles in 9-1-1 loading/unloading. X-ray crystallography (2.1 Å), peptide binding assays, competition assays with RHINO peptide The Journal of biological chemistry High 36841485
2002 Upon replication block (hydroxyurea) in late S phase, co-immunoprecipitation showed interaction of RFC p37 subunit with Rad17, and PCNA with Rad9 and RFC p37. Enhanced colocalization of Rad17 and PCNA was observed in late S phase after hydroxyurea treatment, suggesting Rad17/RFC is recruited to DNA lesions and enables 9-1-1 to interact with PCNA. Co-immunoprecipitation from hydroxyurea-treated HeLa cells, immunofluorescence colocalization Oncogene Low 12400013
2014 In chicken DT40 cells, Rad17 deletion dramatically reduces gene targeting (homologous recombination) efficiency and reduces sister chromatid exchange frequency even in BLM-deficient cells, indicating Rad17 plays a direct role in homologous recombination beyond replication checkpoint signaling. Gene targeting (HR efficiency assay) in Rad17-/- and blm/rad17 DT40 cells, sister chromatid exchange assay, growth curve analysis Genes & genetic systems Medium 19168994
2016 Pol κ depletion in glioblastoma cells facilitates temozolomide-induced Rad17 ubiquitination and proteasomal degradation, thereby silencing ATR-Chk1 signaling. Overexpression of Rad17 in Pol κ-depleted cells restored HR efficiency and desensitized cells to temozolomide, placing Rad17 downstream of Pol κ in protection of stalled replication forks. Ubiquitination assays, western blotting for Rad17 protein levels, HR repair assay, Rad17 overexpression rescue, siRNA knockdown Cancer research Medium 26960975
2014 The Ddc1-Mec3-Rad17 (9-1-1) complex in yeast regulates DNA replication-coupled nucleosome assembly. Rad17 deletion reduces deposition of newly synthesized H3-H4 onto replicated DNA, increases association of histone chaperone Asf1 with Rad53 (reducing Asf1-H3 interaction), and increases H3-H4 interactions with CAF-1 and Rtt106. Genetic epistasis, chromatin immunoprecipitation of newly synthesized histones (SNAP-ChIP), co-immunoprecipitation of Asf1-Rad53 and histone-chaperone interactions in yeast The Journal of biological chemistry Medium 24573675
2014 v-Src suppresses ATR-Chk1 signaling by inhibiting the interaction between Rad17 and Rad9 in the chromatin fraction, without affecting RPA32 phosphorylation, ATR autophosphorylation, or TopBP1-Rad9 interaction, and induces replication fork collapse. Co-immunoprecipitation from chromatin fraction, Chk1 phosphorylation assay, replication fork collapse assay, v-Src overexpression Biochemical and biophysical research communications Low 24971543

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2002 Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin. Genes & development 422 11799063
2003 Replication protein A-mediated recruitment and activation of Rad17 complexes. Proceedings of the National Academy of Sciences of the United States of America 345 14605214
2001 ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses. Nature 231 11418864
2000 Structure-based predictions of Rad1, Rad9, Hus1 and Rad17 participation in sliding clamp and clamp-loading complexes. Nucleic acids research 217 10871397
2003 Yeast Rad17/Mec3/Ddc1: a sliding clamp for the DNA damage checkpoint. Proceedings of the National Academy of Sciences of the United States of America 213 12604797
1995 Fission yeast rad17: a homologue of budding yeast RAD24 that shares regions of sequence similarity with DNA polymerase accessory proteins. The EMBO journal 178 8846774
2000 A novel Rad24 checkpoint protein complex closely related to replication factor C. Current biology : CB 174 10660302
1997 RAD9, RAD17, and RAD24 are required for S phase regulation in Saccharomyces cerevisiae in response to DNA damage. Genetics 169 9017389
2003 Claspin, a Chk1-regulatory protein, monitors DNA replication on chromatin independently of RPA, ATR, and Rad17. Molecular cell 168 12620222
1998 RAD9 and RAD24 define two additive, interacting branches of the DNA damage checkpoint pathway in budding yeast normally required for Rad53 modification and activation. The EMBO journal 128 9564050
2013 Two distinct modes of ATR activation orchestrated by Rad17 and Nbs1. Cell reports 122 23684611
1999 Saccharomyces cerevisiae checkpoint genes MEC1, RAD17 and RAD24 are required for normal meiotic recombination partner choice. Genetics 119 10511543
2004 Human immunodeficiency virus type 1 Vpr-mediated G2 arrest requires Rad17 and Hus1 and induces nuclear BRCA1 and gamma-H2AX focus formation. Molecular and cellular biology 114 15485898
1998 The Saccharomyces cerevisiae RAD9, RAD17, RAD24 and MEC3 genes are required for tolerating irreparable, ultraviolet-induced DNA damage. Genetics 112 9725831
2006 Rad17 phosphorylation is required for claspin recruitment and Chk1 activation in response to replication stress. Molecular cell 105 16885023
2004 Exo1 and Rad24 differentially regulate generation of ssDNA at telomeres of Saccharomyces cerevisiae cdc13-1 mutants. Genetics 102 15454530
1999 Role of a complex containing Rad17, Mec3, and Ddc1 in the yeast DNA damage checkpoint pathway. Molecular and cellular biology 101 9891048
2001 Chl12 (Ctf18) forms a novel replication factor C-related complex and functions redundantly with Rad24 in the DNA replication checkpoint pathway. Molecular and cellular biology 97 11486023
2002 Structures of the human Rad17-replication factor C and checkpoint Rad 9-1-1 complexes visualized by glycerol spray/low voltage microscopy. The Journal of biological chemistry 93 11907025
2004 Interaction between human MCM7 and Rad17 proteins is required for replication checkpoint signaling. The EMBO journal 92 15538388
1998 Human and mouse homologs of Schizosaccharomyces pombe rad1(+) and Saccharomyces cerevisiae RAD17: linkage to checkpoint control and mammalian meiosis. Genes & development 89 9716408
2002 Clamp and clamp loader structures of the human checkpoint protein complexes, Rad9-1-1 and Rad17-RFC. Genes to cells : devoted to molecular & cellular mechanisms 77 12167163
2014 Rad17 recruits the MRE11-RAD50-NBS1 complex to regulate the cellular response to DNA double-strand breaks. The EMBO journal 73 24534091
1998 Identification of a human homologue of the Schizosaccharomyces pombe rad17+ checkpoint gene. The Journal of biological chemistry 71 9660800
1998 Functional and physical interaction between Rad24 and Rfc5 in the yeast checkpoint pathways. Molecular and cellular biology 69 9710632
2000 Rfc5, in cooperation with rad24, controls DNA damage checkpoints throughout the cell cycle in Saccharomyces cerevisiae. Molecular and cellular biology 67 10913172
2000 Genetic studies with the fission yeast Schizosaccharomyces pombe suggest involvement of wee1, ppa2, and rad24 in induction of cell cycle arrest by human immunodeficiency virus type 1 Vpr. Journal of virology 66 10684278
2003 The mitotic DNA damage checkpoint proteins Rad17 and Rad24 are required for repair of double-strand breaks during meiosis in yeast. Genetics 65 12871899
2003 Genomic instability and endoreduplication triggered by RAD17 deletion. Genes & development 61 12672690
2003 The checkpoint protein Rad24 of Saccharomyces cerevisiae is involved in processing double-strand break ends and in recombination partner choice. Molecular and cellular biology 61 12944484
2002 The 14-3-3 proteins Rad24 and Rad25 negatively regulate Byr2 by affecting its localization in Schizosaccharomyces pombe. Molecular and cellular biology 59 12242289
2001 Phosphorylation of serines 635 and 645 of human Rad17 is cell cycle regulated and is required for G(1)/S checkpoint activation in response to DNA damage. Proceedings of the National Academy of Sciences of the United States of America 59 11687627
2010 Rad17 plays a central role in establishment of the interaction between TopBP1 and the Rad9-Hus1-Rad1 complex at stalled replication forks. Molecular biology of the cell 57 20110345
2004 Mutation of the mouse Rad17 gene leads to embryonic lethality and reveals a role in DNA damage-dependent recombination. The EMBO journal 56 15297881
2016 The Error-Prone DNA Polymerase κ Promotes Temozolomide Resistance in Glioblastoma through Rad17-Dependent Activation of ATR-Chk1 Signaling. Cancer research 53 26960975
1998 cDNA cloning and gene mapping of human homologs for Schizosaccharomyces pombe rad17, rad1, and hus1 and cloning of homologs from mouse, Caenorhabditis elegans, and Drosophila melanogaster. Genomics 43 9878245
1996 Cloning and characterization of RAD17, a gene controlling cell cycle responses to DNA damage in Saccharomyces cerevisiae. Nucleic acids research 43 8649984
2003 The human checkpoint Rad protein Rad17 is chromatin-associated throughout the cell cycle, localizes to DNA replication sites, and interacts with DNA polymerase epsilon. Nucleic acids research 41 14500819
1999 HRad17, a human homologue of the Schizosaccharomyces pombe checkpoint gene rad17, is overexpressed in colon carcinoma. Cancer research 40 10232579
2004 Critical role for chicken Rad17 and Rad9 in the cellular response to DNA damage and stalled DNA replication. Genes to cells : devoted to molecular & cellular mechanisms 38 15066121
2015 Phosphorylation-dependent inhibition of Cdc42 GEF Gef1 by 14-3-3 protein Rad24 spatially regulates Cdc42 GTPase activity and oscillatory dynamics during cell morphogenesis. Molecular biology of the cell 37 26246599
2010 Proteolysis of Rad17 by Cdh1/APC regulates checkpoint termination and recovery from genotoxic stress. The EMBO journal 36 20424596
2020 Nitrogen Removal Performance and Metabolic Pathways Analysis of a Novel Aerobic Denitrifying Halotolerant Pseudomonas balearica strain RAD-17. Microorganisms 35 31906569
2019 The miR-205-5p/BRCA1/RAD17 Axis Promotes Genomic Instability in Head and Neck Squamous Cell Carcinomas. Cancers 34 31514456
2001 Fission yeast Rad17 associates with chromatin in response to aberrant genomic structures. Molecular and cellular biology 34 11313455
2018 Warsaw breakage syndrome DDX11 helicase acts jointly with RAD17 in the repair of bulky lesions and replication through abasic sites. Proceedings of the National Academy of Sciences of the United States of America 33 30061412
2008 ATR and Rad17 collaborate in modulating Rad9 localisation at sites of DNA damage. Journal of cell science 32 19020305
2022 Structure of the human RAD17-RFC clamp loader and 9-1-1 checkpoint clamp bound to a dsDNA-ssDNA junction. Nucleic acids research 30 35819203
2011 Replication factor C is a more effective proliferating cell nuclear antigen (PCNA) opener than the checkpoint clamp loader, Rad24-RFC. The Journal of biological chemistry 30 22115746
2006 Recruitment of ATR-ATRIP, Rad17, and 9-1-1 complexes to DNA damage. Methods in enzymology 29 16793398
2013 Depletion of RAD17 sensitizes pancreatic cancer cells to gemcitabine. Journal of cell science 28 23687379
2014 Ubiquitin-specific peptidase 20 regulates Rad17 stability, checkpoint kinase 1 phosphorylation and DNA repair by homologous recombination. The Journal of biological chemistry 27 24923443
2020 IQGAP3 interacts with Rad17 to recruit the Mre11-Rad50-Nbs1 complex and contributes to radioresistance in lung cancer. Cancer letters 26 32896617
2006 Chk1- and claspin-dependent but ATR/ATM- and Rad17-independent DNA replication checkpoint response in HeLa cells. Cancer research 26 16951182
2002 Colocalization of human Rad17 and PCNA in late S phase of the cell cycle upon replication block. Oncogene 26 12400013
2018 Unusual Isothermal Hysteresis in DNA i-Motif pH Transitions: A Study of the RAD17 Promoter Sequence. Biophysical journal 25 29694860
1987 The RAD24 (= Rs1) gene product of Saccharomyces cerevisiae participates in two different pathways of DNA repair. Genetics 25 3549445
2004 Chromatin association of rad17 is required for an ataxia telangiectasia and rad-related kinase-mediated S-phase checkpoint in response to low-dose ultraviolet radiation. Molecular cancer research : MCR 24 15235112
1999 hRAD17, a structural homolog of the Schizosaccharomyces pombe RAD17 cell cycle checkpoint gene, stimulates p53 accumulation. Oncogene 23 10208430
1999 Human and mouse RAD17 genes: identification, localization, genomic structure and histological expression pattern in normal testis and seminoma. Human genetics 21 10480350
1998 The mammalian Rad24 homologous to yeast Saccharomyces cerevisiae Rad24 and Schizosaccharomyces pombe Rad17 is involved in DNA damage checkpoint. Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research 21 9869296
2010 The Saccharomyces cerevisiae RAD9, RAD17 and RAD24 genes are required for suppression of mutagenic post-replicative repair during chronic DNA damage. DNA repair 19 20472512
2001 Human Rad17 is phosphorylated upon DNA damage and also overexpressed in primary non-small cell lung cancer tissues. Cancer research 18 11606373
2013 Rad9, Rad17, TopBP1 and claspin play essential roles in heat-induced activation of ATR kinase and heat tolerance. PloS one 17 23383325
2009 Identification of sam4 as a rad24 allele in Schizosaccharomyces pombe. Bioscience, biotechnology, and biochemistry 17 19584544
2000 A novel mutant allele of the chromatin-bound fission yeast checkpoint protein Rad17 separates the DNA structure checkpoints. Journal of cell science 17 10683155
2013 Regulation of Rad17 protein turnover unveils an impact of Rad17-APC cascade in breast carcinogenesis and treatment. The Journal of biological chemistry 16 23637229
2006 HIV-1 Vpr induces G2 cell cycle arrest in fission yeast associated with Rad24/14-3-3-dependent, Chk1/Cds1-independent Wee1 upregulation. Microbes and infection 16 16968670
1999 Human and mouse homologs of the Schizosaccharomyces pombe rad17+ cell cycle checkpoint control gene. Genomics 16 9933569
2014 Genetic and physical interaction of Ssp1 CaMKK and Rad24 14-3-3 during low pH and osmotic stress in fission yeast. Open biology 15 24451546
2007 The Caenorhabditis elegans Rad17 homolog HPR-17 is required for telomere replication. Genetics 15 17339221
2005 Function of Rad17/Mec3/Ddc1 and its partial complexes in the DNA damage checkpoint. DNA repair 15 16137930
2001 RAD9, RAD24, RAD16 and RAD26 are required for the inducible nucleotide excision repair of UV-induced cyclobutane pyrimidine dimers from the transcribed and non-transcribed regions of the Saccharomyces cerevisiae MFA2 gene. Mutation research 15 11267834
2012 Abasic sites linked to dUTP incorporation in DNA are a major cause of spontaneous mutations in absence of base excision repair and Rad17-Mec3-Ddc1 (9-1-1) DNA damage checkpoint clamp in Saccharomyces cerevisiae. DNA repair 14 22226374
2016 The KYxxL motif in Rad17 protein is essential for the interaction with the 9-1-1 complex. Biochemical and biophysical research communications 13 27387238
2022 Genetic screen for suppression of transcriptional interference reveals fission yeast 14-3-3 protein Rad24 as an antagonist of precocious Pol2 transcription termination. Nucleic acids research 12 34967420
2015 Chemogenetic profiling identifies RAD17 as synthetically lethal with checkpoint kinase inhibition. Oncotarget 12 26437225
2008 Downregulation of RAD17 in head and neck cancer. Head & neck 12 17657792
2002 A dominant-negative MEC3 mutant uncovers new functions for the Rad17 complex and Tel1. Proceedings of the National Academy of Sciences of the United States of America 11 12271137
2019 Human Rad17 C-terminal tail is phosphorylated by concerted action of CK1δ/ε and CK2 to promote interaction with the 9-1-1 complex. Biochemical and biophysical research communications 9 31353086
2019 Distinct Functions in Regulation of Meiotic Crossovers for DNA Damage Response Clamp Loader Rad24(Rad17) and Mec1(ATR) Kinase. Genetics 9 31597673
2014 v-Src inhibits the interaction between Rad17 and Rad9 and induces replication fork collapse. Biochemical and biophysical research communications 9 24971543
2000 Rad24 is essential for proliferation of diploid cells in fission yeast. FEBS letters 9 10788621
2022 Impairment of RAD17 Functions by miR-506-3p as a Novel Synthetic Lethal Approach Targeting DNA Repair Pathways in Ovarian Cancer. Frontiers in oncology 8 35924161
2010 Roles of the checkpoint sensor clamp Rad9-Rad1-Hus1 (911)-complex and the clamp loaders Rad17-RFC and Ctf18-RFC in Schizosaccharomyces pombe telomere maintenance. Cell cycle (Georgetown, Tex.) 8 20505337
2023 The 9-1-1 DNA clamp subunit RAD1 forms specific interactions with clamp loader RAD17, revealing functional implications for binding-protein RHINO. The Journal of biological chemistry 7 36841485
2021 Performance of Aerobic Denitrification by the Strain Pseudomonas balearica RAD-17 in the Presence of Antibiotics. Microorganisms 6 34442663
2018 Casein kinase 2 promotes interaction between Rad17 and the 9-1-1 complex through constitutive phosphorylation of the C-terminal tail of human Rad17. Biochemical and biophysical research communications 6 29902452
2017 The polyanionic C-terminal tail of human Rad17 regulates interaction with the 9-1-1 complex. Biochemical and biophysical research communications 6 28666868
2015 Characterization of the interaction between Rfa1 and Rad24 in Saccharomyces cerevisiae. PloS one 6 25719602
2007 Human Raf-1 proteins associate with Rad24 and Cdc25 in cell-cycle checkpoint pathway of fission yeast, Schizosaccharomyces pombe. Journal of cellular biochemistry 6 17243098
2003 Correlation between checkpoint activation and in vivo assembly of the yeast checkpoint complex Rad17-Mec3-Ddc1. The Journal of biological chemistry 6 12672803
2001 Multiple alternative splicing forms of human RAD17 and their differential response to ionizing radiation. Gene 6 11602352
2019 The structure of the checkpoint clamp 9-1-1 complex and clamp loader Rad24-RFC in Saccharomyces cerevisiae. Biochemical and biophysical research communications 5 31182279
2017 The fission yeast MAPK Spc1 senses perturbations in Cdc25 and Wee1 activities and targets Rad24 to restore this balance. Yeast (Chichester, England) 5 29065217
2008 A novel role for Rad17 in homologous recombination. Genes & genetic systems 5 19168994
2020 The noncoding function of NELFA mRNA promotes the development of oesophageal squamous cell carcinoma by regulating the Rad17-RFC2-5 complex. Molecular oncology 4 31845510
2014 The Ddc1-Mec3-Rad17 sliding clamp regulates histone-histone chaperone interactions and DNA replication-coupled nucleosome assembly in budding yeast. The Journal of biological chemistry 4 24573675
2008 The human homolog of fission yeast Rad17 is implicated in tumor growth. Cancer letters 4 18378394
2008 HDF1 and RAD17 genes are involved in DNA double-strand break repair in stationary phase Saccharomyces cerevisiae. Journal of biological physics 4 19669493

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