Affinage

RAD50

DNA repair protein RAD50 · UniProt Q92878

Length
1312 aa
Mass
153.9 kDa
Annotated
2026-06-10
100 papers in source corpus 47 papers cited in narrative 45 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

RAD50 is the ABC-ATPase scaffold subunit of the conserved MRE11-RAD50-NBS1 (MRN) complex, the primary cellular sensor and processor of DNA double-strand breaks (DSBs) (PMID:8756642, PMID:9590181, PMID:10346816). RAD50 stably associates with MRE11 to form a (MRE11)2(RAD50)2 heterotetrameric DNA-processing head linked to long coiled-coil arms, with NBS1 completing the trimeric complex required for ionizing-radiation-induced nuclear foci and DSB-checkpoint coupling (PMID:9590181, PMID:11371344). RAD50 supplies the complex's ATP-binding and hydrolysis activity, which acts as a conformational switch: ATP binding drives a 'closed' state that promotes DNA-end binding, end tethering, and ATM/Tel1 kinase activation, while ATP hydrolysis opens the complex to unmask the MRE11 active site and convert it from endonuclease to exonuclease, enabling endonucleolytic incision past protein adducts and Ku blocks followed by 5'-strand resection (PMID:10346816, PMID:21511873, PMID:22102415, PMID:24493214, PMID:26717941). The coiled-coil arms terminate in a Zn2+-coordinated zinc-hook dimerization interface that bridges and tethers broken DNA ends across long distances, and conformational changes are transmitted from the hook through the coiled coils to the globular ATPase/nuclease domain (PMID:12152085, PMID:16163361, PMID:25601756). Both RAD50 ATPase active sites are required for DNA-stimulated ATP hydrolysis, adduct cleavage, and ATM activation, and the complex stimulates ATM by tethering DNA and recruiting downstream factors including CtIP and EXO1 to drive resection (PMID:28867292, PMID:28369545, PMID:15064416, PMID:16622404). MRN directly activates ATM toward p53, Chk2, and H2AX, and RAD50 is itself phosphorylated by ATM at Ser-635 to act as an adaptor for downstream checkpoint signaling through SMC1 (PMID:15064416, PMID:21757780). Beyond DSB repair, RAD50 participates in telomere length control via MRN/ATM regulation of TRF1 (PMID:17694070) and, in the cytoplasm, interacts with CARD9 to couple cytosolic dsDNA sensing to NF-κB activation and pro-IL-1β production (PMID:24777530).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 1996 High

    Established that human RAD50 is the homolog of yeast Rad50 and acts within a multiprotein complex rather than alone, defining the molecular context for all subsequent mechanism.

    Evidence Co-immunoprecipitation and cDNA characterization identifying stable RAD50-MRE11 association

    PMID:8756642

    Open questions at the time
    • Did not define the third subunit
    • No enzymatic or structural mechanism
  2. 1998 High

    Identified NBS1 as the third MRN subunit and connected the complex to a human genome-instability disease, linking DSB repair to checkpoint signaling.

    Evidence Gene mapping and immunofluorescence in NBS patient cells showing NBS1 dependence of RAD50/MRE11 IR foci

    PMID:9315668 PMID:9590181

    Open questions at the time
    • Mechanism of foci assembly not resolved
    • Catalytic roles of each subunit unknown
  3. 1999 High

    Assigned enzymatic functions within the reconstituted complex, showing RAD50 provides ATP binding that switches MRE11 nuclease specificity.

    Evidence Biochemical reconstitution with recombinant NBS1/MRE11/RAD50, nuclease and helicase assays with mutants

    PMID:10346816 PMID:11454871

    Open questions at the time
    • Structural basis of the ATP switch not yet visualized
    • In vivo relevance of ATP-controlled specificity not tested
  4. 2002 High

    Resolved the structural architecture: RAD50 is an ABC-ATPase whose coiled coils end in a Zn2+ zinc-hook dimer interface enabling long-range DNA tethering.

    Evidence X-ray crystallography of archaeal Rad50/Mre11, EM, and yeast genetics of zinc-hook mutants

    PMID:11371344 PMID:12152085 PMID:14698290

    Open questions at the time
    • How hook signal propagates to the catalytic head not yet shown
    • Eukaryotic full-complex architecture unresolved
  5. 2004 High

    Demonstrated that MRN directly activates ATM kinase, establishing RAD50's complex as an upstream signaling activator and not merely a repair enzyme.

    Evidence In vitro kinase assays with recombinant MRN and ATM substrates p53, Chk2, H2AX

    PMID:15064416

    Open questions at the time
    • Did not separate tethering from direct activation contributions
    • RAD50-specific contribution to ATM activation not isolated
  6. 2005 High

    Visualized the DNA-induced conformational change that converts intracomplex coiled-coil interactions to intercomplex tethering, providing a physical mechanism for end bridging.

    Evidence Atomic force and electron microscopy and single-molecule imaging of hRAD50/MRE11/NBS1 on DNA

    PMID:16163361

    Open questions at the time
    • Coupling to ATP cycle quantitative detail incomplete
    • Nuclease activation step not captured
  7. 2006 High

    Clarified that MRN promotes ATM activation in two steps, with RAD50-mediated DNA tethering increasing local ATM concentration to drive monomer conversion.

    Evidence Reconstitution in Xenopus cell-free extracts with NBS1 ATM-binding-domain mutants

    PMID:16622404

    Open questions at the time
    • RAD50 ATPase-state requirement not directly tested here
    • In-cell ordering of steps not established
  8. 2011 High

    Defined the ATP-driven molecular switch at atomic resolution and tied RAD50 Ser-635 ATM phosphorylation to downstream checkpoint signaling.

    Evidence Mre11-Rad50-ATPγS crystal structure with biochemistry; phosphosite mapping and S635G complementation in RAD50-deficient cells

    PMID:21511873 PMID:21757780 PMID:22102415

    Open questions at the time
    • Kinetics of the endo-to-exo transition in vivo not measured
    • S635 effector pathway beyond SMC1 incompletely mapped
  9. 2013 High

    Showed RAD50 ATPase activity directly powers DNA-end unwinding required for resection, linking the catalytic cycle to repair output in human cells.

    Evidence Single-molecule FRET unwinding assays with RAD50 catalytic mutants plus cellular repair assays

    PMID:24191051

    Open questions at the time
    • Coupling of unwinding to nucleolytic incision not fully resolved
    • Role of accessory factors during unwinding not addressed
  10. 2014 High

    Established the structural separation between the closed-state tethering/signaling function and hydrolysis-driven opening required for resection, and revealed a cytoplasmic RAD50-CARD9 innate-immune role.

    Evidence Structure-based Rad50 mutants across organisms; RAD50-CARD9 Co-IP and conditional-knockout cytokine and infection assays

    PMID:24493214 PMID:24694934 PMID:24777530

    Open questions at the time
    • Structural basis of RAD50-CARD9 interaction unknown
    • Crosstalk between nuclear repair and cytoplasmic immune roles unexplored
  11. 2017 High

    Demonstrated that both RAD50 ATPase sites are jointly required for adduct cleavage and ATM activation, and that RAD50 enables MRN to translocate on DNA and load EXO1 for processive resection.

    Evidence Single-active-site Rad50 dimer mutants in ATPase/nuclease/ATM assays; single-molecule microscopy of MRN-EXO1 on DNA

    PMID:28134932 PMID:28369545 PMID:28867292

    Open questions at the time
    • Coordination of the two active sites in vivo not directly observed
    • Adduct-specificity determinants incompletely defined
  12. 2019 High

    Resolved the conformational mechanics of DNA clamping and nuclease loading and identified C1QBP as a phosphorylation-controlled negative regulator of MRE11/RAD50 nuclease.

    Evidence Cryo-EM of SbcCD resting and DNA-bound states; Co-IP, nuclease, and ATM-phosphosite analysis of the C1QBP-MRE11-RAD50 complex

    PMID:30819891 PMID:31353207 PMID:31492634

    Open questions at the time
    • Eukaryotic full-complex conformational cycle not yet at high resolution
    • Regulation of C1QBP dissociation timing in cells incomplete
  13. 2022 High

    Delivered near-complete eukaryotic MRN architecture and RAD50-mediated higher-order oligomerization, explaining how RAD50 orients MRE11 for distinct endo- versus exonucleolytic cuts and bridges paired complexes.

    Evidence Cryo-EM of C. thermophilum MRN and SbcCD on diverse substrates; EM and yeast genetics of Rad50 oligomerization mutants

    PMID:35501303 PMID:35987200 PMID:36577401

    Open questions at the time
    • Functional role of MRN-MRN dimerization in cells incompletely defined
    • Mechanism converting orientation to substrate selection not fully resolved

Open questions

Synthesis pass · forward-looking unresolved questions
  • How RAD50's nuclear DSB-processing functions are coordinated with its cytoplasmic innate-immune signaling role, and the structural basis of RAD50-CARD9 dsDNA sensing, remain open.
  • No structure of the dsDNA-RAD50-CARD9 complex
  • No mechanism partitioning RAD50 between nucleus and cytoplasm
  • Disease-causing RAD50 mutation effects on either role not characterized in this corpus

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140657 ATP-dependent activity 6 GO:0016787 hydrolase activity 5 GO:0140097 catalytic activity, acting on DNA 5 GO:0005198 structural molecule activity 4 GO:0003677 DNA binding 3 GO:0016740 transferase activity 1
Localization
GO:0005634 nucleus 3 GO:0000228 nuclear chromosome 2 GO:0005730 nucleolus 2 GO:0005829 cytosol 1
Pathway
R-HSA-73894 DNA Repair 5 R-HSA-8953897 Cellular responses to stimuli 4 R-HSA-1640170 Cell Cycle 2 R-HSA-1852241 Organelle biogenesis and maintenance 2 R-HSA-168256 Immune System 1
Partners
BRCA1C1QBPCARD9CTIPEXO1MRE11NBS1RINT-1
Complex memberships
MRC complex (MRE11-RAD50-C1QBP)MRN complex (MRE11-RAD50-NBS1)MRX complex (Mre11-Rad50-Xrs2)

Evidence

Reading pass · 45 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1996 Human RAD50 protein is stably associated with human MRE11 in a protein complex, identified as the human homolog of yeast Rad50 which functions in a multiprotein complex analogous to the yeast Rad50/Mre11/Xrs2 complex. Co-immunoprecipitation, cDNA isolation and characterization Molecular and cellular biology High 8756642
1997 hMRE11 and hRAD50 form discrete nuclear foci in response to DNA double-strand break-inducing agents (ionizing radiation) but not UV irradiation, and focus formation is markedly reduced in ataxia-telangiectasia cells, indicating dependence on a DNA damage-induced signaling pathway. Immunofluorescence, cell irradiation experiments Molecular and cellular biology High 9315668
1998 The hMRE11/hRAD50 complex includes a third component, p95/NBS1 (the gene mutated in Nijmegen breakage syndrome); p95 deficiency abrogates formation of hMRE11/hRAD50 ionizing radiation-induced foci, linking DSB repair to cell cycle checkpoint functions. Gene mapping, immunofluorescence, cell biology in NBS patient-derived cells Cell High 9590181
1999 BRCA1 interacts in vitro and in vivo with hRAD50 (which forms a complex with hMRE11 and p95/nibrin); upon irradiation, BRCA1 co-localizes with hRAD50 in nuclear foci, and wild-type BRCA1 is required for formation of these foci and for cellular resistance to DNA damage. In vitro binding assay, co-immunoprecipitation, immunofluorescence colocalization Science High 10426999
1999 The triple complex of recombinant NBS1, MRE11, and RAD50 displays ATP-stimulated DNA unwinding and efficient cleavage of fully paired hairpins not seen with MRE11/RAD50 alone; RAD50 is responsible for ATP binding by the complex, and ATP controls a switch in endonuclease specificity allowing cleavage of 3'-protruding strands at double-/single-strand transitions. Biochemical reconstitution with recombinant proteins, mutational analysis, in vitro nuclease/helicase assays Genes & development High 10346816
2000 A small fraction of RAD50, MRE11, and NBS1 is associated with TRF2 immunocomplexes; RAD50 and MRE11 localize to interphase telomeres, and NBS1 associates with TRF2 and telomeres specifically in S phase but not G1 or G2, indicating cell-cycle-regulated association of MRN with telomeres. Nanoelectrospray tandem mass spectrometry, protein blotting, indirect immunofluorescence Nature genetics High 10888888
2001 Crystal structures of Pyrococcus furiosus Mre11 reveal a protein phosphatase-like dimanganese-binding nuclease domain; the structure of P. furiosus Rad50 ABC-ATPase with adjacent coiled-coil defines a compact Mre11/Rad50-ATPase complex and suggests that Rad50 ATP-driven conformational switching directly controls the Mre11 exonuclease; the MR complex exists as a (Mre11)2/(Rad50)2 heterotetrameric DNA-processing head with a double coiled-coil linker. X-ray crystallography, electron microscopy, small-angle X-ray scattering, ultracentrifugation Cell High 11371344
2001 Purified yeast Rad50 and Mre11 form a stable equimolar complex. Mre11 has 3'→5' exonuclease activity releasing mononucleotides; addition of Rad50 does not significantly alter this exonuclease activity. Mre11 has endonuclease activity on hairpins and 3' ssDNA tails, and these endonuclease activities are markedly enhanced by Rad50 only in the presence of ATP. Protein purification, in vitro nuclease assays, ATP-dependence studies The Journal of biological chemistry High 11454871
2001 NBS1 C-terminal sequence (residues 665-693) is essential for hMRE11 binding and is necessary for nuclear localization of the MRE11/RAD50 complex and cellular radiation resistance; the N-terminal FHA domain regulates nuclear foci formation in response to DNA damage but is not essential for nuclear transport or radiation resistance. Deletion mutagenesis, co-immunoprecipitation, cellular localization experiments, radiation sensitivity assays The Journal of biological chemistry High 11062235
2002 The Rad50 coiled-coil region contains a zinc-hook dimer interface where pairs of conserved Cys-X-X-Cys motifs form interlocking hooks that bind one Zn2+ ion; these hooks join oppositely protruding Rad50 coiled-coil domains to form a flexible bridge of up to 1,200 Å, enabling DNA tethering. Mutations in this zinc-hook motif confer radiation sensitivity in yeast and disrupt binding at the distant Mre11 nuclease interface. X-ray crystallography (2.2 Å), electron microscopy, biochemical assays, yeast genetics Nature High 12152085
2002 Rad50 ATP-binding domains share structural and mechanistic conservation with ABC transporters; ATP binding drives conformational changes in substrate-specific domains, functioning as a chemo-mechanical device in DNA repair. Structural analysis, comparative crystallography Current opinion in structural biology Medium 12727520
2002 Rad50 S793R signature motif mutation prevents ATP binding and disrupts communication among other ATP-binding loops, prevents Rad50 dimerization; the equivalent mutation in human RAD50 forms a complex with MRE11 and NBS1 but is specifically deficient in all ATP-dependent enzymatic activities; the same mutation in S. cerevisiae fails to complement rad50 deletion in DNA repair assays. X-ray crystallography, biochemical ATPase assays, yeast genetic complementation Journal of molecular biology High 14698290
2004 The MRN complex directly activates ATM kinase activity in vitro toward substrates p53, Chk2, and histone H2AX; MRN makes multiple contacts with ATM and stimulates ATM activity by facilitating stable substrate binding; phosphorylation of NBS1 is critical for MRN stimulation of ATM activity toward Chk2 but not p53. In vitro kinase assays with recombinant proteins, protein interaction studies Science High 15064416
2004 A novel RAD50-interacting protein, RINT-1, was identified via yeast two-hybrid screening; RINT-1 binds specifically to RAD50 during late S and G2/M phases of the cell cycle; cells expressing truncated RINT-1 display a defective radiation-induced G2/M checkpoint. Yeast two-hybrid screen, co-immunoprecipitation, cell cycle analysis, checkpoint assay The Journal of biological chemistry Medium 11096100
2004 RPA and the MRN complex (MRE11, RAD50, NBS1) co-localize to discrete nuclear foci and physically interact in response to DNA replication fork blockage by HU or UV; co-immunoprecipitation of RPA with anti-RAD50 antibody was demonstrated; phosphorylation of both RPA and MRE11 is required for this interaction. Co-immunoprecipitation, immunofluorescence, subcellular fractionation, phosphatase treatment The Journal of biological chemistry Medium 15180989
2005 DNA binding by the hRAD50/MRE11/NBS1 globular domain leads to parallel orientation of the coiled coils, preventing intracomplex interactions and favoring intercomplex associations needed for DNA tethering; this conformational change is transmitted 50 nm from the DNA-binding domain to the coiled-coil apices. Atomic force microscopy, electron microscopy, single-molecule imaging Nature High 16163361
2006 ATM activation by DSBs occurs in two steps: first, dimeric ATM is recruited to damaged DNA and dissociates into monomers (facilitated by MRN tethering DNA to increase local concentration); second, the ATM-binding domain of NBS1 is required and sufficient to convert unphosphorylated ATM monomers into enzymatically active monomers in the absence of DNA. Biochemical reconstitution, Xenopus cell-free extract system, mutational analysis Nature structural & molecular biology High 16622404
2007 MRE11-RAD50-NBS1 and ATM function as co-mediators of TRF1 in telomere length control; RAD50 targeted to telomeres downregulates TRF1 association with telomeric DNA; ATM-mediated phosphorylation of TRF1 impairs its ability to bind telomeric DNA in vitro, and MRN is required for this TRF1 phosphorylation by ATM. Chromatin immunoprecipitation, telomere-targeted overexpression, in vitro DNA binding assays, RNAi knockdown Nature structural & molecular biology Medium 17694070
2007 Mre11/Rad50 complexes from three organisms (human, yeast, archaeal) catalyze the reversible adenylate kinase reaction in vitro; mutation of the conserved signature motif reduces adenylate kinase activity without reducing ATP hydrolysis; this mutant resembles a rad50 null strain for meiosis and telomere maintenance; adenylate kinase inhibitor blocks Mre11/Rad50-dependent DNA tethering in vitro and in cell-free extracts. In vitro enzymatic assays, yeast genetics, pharmacological inhibition, cell-free extract DNA tethering assay Molecular cell High 17349953
2009 NBS1 FHA domain recruits phosphorylated Ctp1 to DSBs via binding of the NBS1 FHA domain to a Ctp1 pThr-Asp motif; fission yeast and human NBS1 structures reveal fused FHA-BRCT1-BRCT2 domains flexibly linked to C-terminal Mre11- and ATM-binding motifs; Nbs1 tethers Ctp1/CtIP to the immediate vicinity of DSBs to restrict DNA end processing activities. X-ray crystallography, SAXS, genetic analysis, biochemical binding assays Cell High 19804755
2010 Biochemical reconstitution of DNA end resection shows that the yeast MRX complex and Sae2 stimulate Exo1-mediated 5' strand degradation through cooperative binding of DNA substrates; MRX recruits Dna2 nuclease to DSB ends and stimulates recruitment of Exo1 while antagonizing excess Ku binding to DSB ends. In vitro resection reconstitution with purified proteins, ChIP in yeast Nature; The EMBO journal High 20811461 20834227
2010 MRX and Sae2 directly promote 5' strand resection; reconstitution with purified MRX, Sae2, and Exo1 shows that 5' strand degradation is catalyzed by Exo1 yet completely dependent on MRX and Sae2 when Exo1 levels are limiting, mainly through cooperative DNA substrate binding. In vitro resection reconstitution with purified recombinant proteins Nature structural & molecular biology High 21102445
2011 Crystal structure of the Mre11-Rad50-ATPγS complex shows Mre11 promotes ATPase activity of Rad50 by holding the coiled-coil arm and stabilizing the signature motif and P-loop; ATP-bound Rad50 negatively regulates Mre11 nuclease activity by blocking the Mre11 active site; ATP hydrolysis disengages Rad50 molecules and causes conformational change in Mre11's flexible linker to unmask the Mre11 active site. X-ray crystallography, biochemical ATPase and nuclease assays Genes & development High 21511873
2011 ATM phosphorylates RAD50 at Ser-635; a RAD50 S635G phosphosite mutant supports normal ATM activation but is defective in correcting DNA damage-induced signaling through ATM-dependent substrate SMC1, fails to correct radiosensitivity, DSB repair, and S-phase checkpoint defects, without disrupting MRE11/RAD50/NBS1 complex integrity. Phosphorylation site mapping, site-directed mutagenesis, functional complementation in RAD50-deficient cells The Journal of biological chemistry High 21757780
2011 ATP binding to RAD50 induces a closed conformation in which MRE11 functions as an endonuclease; ATP hydrolysis opens the RAD50-MRE11 complex and MRE11 maintains exonuclease activity. Thus, ATP hydrolysis acts as a molecular switch converting MRE11 from endonuclease to exonuclease. In vitro nuclease assays, EM visualization of conformational states, ATP binding/hydrolysis studies The Journal of biological chemistry High 22102415
2013 MRN complex visualized by single-molecule FRET unwinds 15-20 base pairs at the end of a duplex DNA molecule, holding the branched structure open for minutes in an ATP-dependent reaction; a Rad50 catalytic domain mutant deficient in this ATP-dependent DNA unwinding is impaired in DNA end resection in vitro and in resection-dependent repair in human cells. Single-molecule FRET, in vitro resection assays, human cell DNA repair assays Proceedings of the National Academy of Sciences High 24191051
2014 RAD50 interacts directly with innate immune adaptor CARD9 in the cytoplasm; dsDNA induces formation of dsDNA-RAD50-CARD9 signaling complexes activating NF-κB and generating pro-IL-1β; cells conditionally deficient in RAD50 exhibit defective DNA-induced IL-1β production. Co-immunoprecipitation, conditional knockout cells, cytokine assays, in vivo infection model Nature immunology High 24777530
2014 Structure-based mutations in Rad50 that promote or destabilize the ATP-bound 'closed' state show that the closed conformation promotes DNA end binding and end tethering, while hydrolysis-induced opening is essential for DNA resection; reducing ATP-bound state stability impairs DNA repair and Tel1/ATM checkpoint signaling in S. pombe, DSB resection in S. cerevisiae, and ATM activation by human MRN in vitro. Crystal structures, X-ray scattering, biochemical assays, genetic functional analyses in multiple organisms The EMBO journal High 24493214
2014 RAD50 phosphorylation at S635 by ATR is required for ATR signaling through Chk1 and downstream substrates in response to replication stress; RAD50 S635 phosphorylation is essential for DNA replication restart by promoting cohesin loading at stalled replication forks; RAD50 is required for ATR activation in mammalian cells. Phosphorylation site mutagenesis, functional complementation, replication restart assay, co-immunoprecipitation Human molecular genetics Medium 24694934
2015 Crystal structure of Methanococcus jannaschii MR-ATPγS-DNA shows that partly deformed DNA runs symmetrically across the central groove between two ATPγS-bound Rad50 nucleotide-binding domains; duplex DNA cannot access the Mre11 active site in the ATP-free full-length MR complex; ATP hydrolysis drives rotation of the nucleotide-binding domain inducing DNA melting so substrate DNA can access Mre11. X-ray crystallography, in vitro biochemical assays The EMBO journal High 26717941
2015 Rad50 hook mutations flanking Zn2+-coordinating cysteines impair hook-mediated dimerization; Mre11 complex functions specified by the globular domain including Tel1/ATM activation, NHEJ, and DSB end resection are affected; these phenotypes are suppressed by mutations within the coiled-coil and globular ATPase domains, indicating conformational changes are transmitted from the hook through the coiled coils to the globular domain. Yeast genetics, biochemical assays, suppressor analysis Molecular cell High 25601756
2016 Human MRN complex (hMRN) catalyzes sequential endonucleolytic and exonucleolytic activities on both 5' and 3' strands of DNA ends containing protein adducts; NBS1, ATP, and adducts are essential for this function; NBS1 inhibits MRE11/RAD50-catalyzed 3'→5' exonucleolytic degradation of clean DNA ends; phosphorylated CtIP further stimulates hMRN endonucleolytic cleavage. In vitro nuclease assays with recombinant proteins, mutational analysis Molecular cell High 27814491
2016 Crystal structure of ATP-bound eukaryotic Rad50 NBD dimer (Chaetomium thermophilum) in complex with DNA shows that a Rad50 dimer binds approximately 18 bp of DNA along the dimer interface in an ATP-dependent fashion, or bridges two DNA ends with a preference for 3' overhangs; a strand-loop-helix motif on Rad50 NBD mediates DNA binding. X-ray crystallography, SAXS, cross-linking studies, in vitro DNA binding, yeast mutational analysis The EMBO journal High 26896444
2017 RAD50 binds homoduplex DNA and promotes one-dimensional facilitated diffusion of MRN along DNA including nucleosome-coated DNA; MRE11 is required for DNA end recognition and nuclease activities; MRN removes Ku or other DNA adducts via an MRE11-dependent nucleolytic reaction; MRN then loads EXO1 onto free DNA ends and acts as a processivity factor for EXO1 in the presence of RPA. High-throughput single-molecule microscopy, in vitro reconstitution with purified proteins Molecular cell High 28867292
2017 Human Rad50 hook and coiled-coil domain structure reveals a predominant intracomplex rod-shaped dimer in which the two parallel coiled coils form a rod; a novel interface within the coiled-coil domains stabilizes the Rad50 protomer interaction; in yeast, removal of this coiled-coil interface compromises Tel1/ATM activation without affecting DNA repair. X-ray crystallography, yeast genetic analysis Nature structural & molecular biology High 28134932
2017 Both ATPase active sites of the Rad50 dimer must be functional for stimulation of ATP hydrolysis by DNA ends, for endonucleolytic cleavage at protein adducts, and for stimulation of ATM kinase activity; double-stranded DNA stimulates ATP hydrolysis by hMRN ~20-fold in an end-dependent manner. In vitro ATPase assays with catalytic site mutants creating single-active-site Rad50 dimers, in vitro nuclease and ATM kinase assays Nucleic acids research High 28369545
2018 The Ku complex shields DNA ends from MRX exonuclease activity but facilitates MRX endonucleolytic scission of the 5'-terminated strand in an ATP- and Sae2-dependent manner; the endonucleolytic incision site is enlarged into a gap via MRX exonuclease activity stimulated by Sae2 without ATP; RPA renders partially resected DNA susceptible to MRX-Sae2; internal protein blocks trigger DNA cleavage by MRX. In vitro resection assays with purified proteins, yeast genetics Genes & development High 29321177 29321179
2019 Cryo-EM structures of bacterial Mre11-Rad50 homolog SbcCD in resting and DNA-bound states reveal: in resting state Mre11 nuclease is blocked by ATP-Rad50; upon DNA binding, the two coiled coils zip up into a rod forming a clamp around dsDNA; Mre11 moves to the side of Rad50, binds the DNA end, and assembles a DNA cutting channel for nuclease reactions. Cryo-electron microscopy, biochemical validation Molecular cell High 31492634
2019 The ATP-bound 'closed' conformation of the Mre11-Rad50 complex is essential for Tel1/ATM activation; separation-of-function mutants mre11-S499P and rad50-A78T specifically reduce Tel1-MRX interaction and Tel1 association at DSBs; Rad50-A78T destabilizes the ATP-bound conformational state; molecular dynamics simulations confirm that ATP-bound MR complex lingers in a tightly closed conformation. Yeast genetics, biochemical Tel1 kinase assays, molecular dynamics simulations, ChIP Nucleic acids research High 30698745
2019 Stepwise resection by MRX-Sae2 proceeds through endonucleolytic DNA incisions followed by exonucleolytic 3'→5' degradation of individual DNA fragments; Rad50 restricts Mre11 exonuclease in an ATP binding-dependent manner preventing 3' end degradation; phosphorylated Sae2 overcomes this inhibition to promote MRX 3'→5' exonuclease activity, which requires ATP hydrolysis by Rad50. In vitro resection assays with purified recombinant proteins and plasmid-length substrates Proceedings of the National Academy of Sciences High 30819891
2019 C1QBP forms a complex (MRC) with MRE11 and RAD50, stabilizing MRE11/RAD50 while inhibiting MRE11 nuclease activity by preventing DNA/chromatin binding; upon DNA damage, ATM phosphorylates MRE11-S676/S678 to dissociate the MRC complex, allowing MRN recruitment to DSBs. Co-immunoprecipitation, in vitro nuclease assays, mutagenesis, chromatin fractionation Molecular cell High 31353207
2019 Tel1/ATM activation by MRX requires Rad50 ATPase activity and long nucleosome-free dsDNA but not DNA ends; either Mre11 or Xrs2 (but not necessarily both) is required in addition to DNA and Rad50; all three MRX subunits physically associate with Tel1. In vitro Tel1 kinase assays with purified components, co-immunoprecipitation, systematic mutant analysis The Journal of biological chemistry High 31073030
2022 Cryo-EM structure of Chaetomium thermophilum MRN (2:2:1 complex) reveals: single NBS1 wrapping around the autoinhibited Mre11 nuclease dimer; two DNA-binding modes (ATP-dependent for DNA ends, ATP-independent through Mre11 C-terminus); two 60-nm coiled-coil domains forming a linear rod with zinc-hook apices; two MRN complexes can dimerize via hook apices to form 120-nm spanning MRN-MRN structures. Cryo-electron microscopy, biochemical validation Molecular cell High 36577401
2022 Cryo-EM structures of bacterial SbcCD (Mre11-Rad50 homolog) bound to protein-blocked DNA end and DNA hairpin show: Mre11-Rad50 bends internal DNA for endonucleolytic cleavage; Mre11 is loaded onto blocked DNA ends with Mre11 pointing away from the block, explaining how 3'→5' exonuclease and endonuclease activities are mechanistically distinguished by the orientation of Mre11-Rad50 on diverse DNA ends. Cryo-electron microscopy, biochemical nuclease assays Molecular cell High 35987200
2022 S. cerevisiae Mre11-Rad50 (with or without Xrs2) forms higher-order oligomeric assemblies in solution and on DNA; Rad50 mediates oligomerization through a conserved beta-sheet; mutations disrupting oligomerization impair foci formation, DNA damage signaling, DSB repair, and telomere maintenance in vivo; oligomerization does not affect exonuclease activity but drives endonucleolytic cleavage at multiple sites on the 5'-DNA strand. Electron microscopy, biochemical assays, yeast genetic analysis, in vitro nuclease assays Nature communications High 35501303

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1998 The hMre11/hRad50 protein complex and Nijmegen breakage syndrome: linkage of double-strand break repair to the cellular DNA damage response. Cell 986 9590181
2004 Direct activation of the ATM protein kinase by the Mre11/Rad50/Nbs1 complex. Science (New York, N.Y.) 602 15064416
1999 Association of BRCA1 with the hRad50-hMre11-p95 complex and the DNA damage response. Science (New York, N.Y.) 513 10426999
2000 Cell-cycle-regulated association of RAD50/MRE11/NBS1 with TRF2 and human telomeres. Nature genetics 491 10888888
2002 The Rad50 zinc-hook is a structure joining Mre11 complexes in DNA recombination and repair. Nature 437 12152085
1999 Nbs1 potentiates ATP-driven DNA unwinding and endonuclease cleavage by the Mre11/Rad50 complex. Genes & development 434 10346816
2001 Structural biochemistry and interaction architecture of the DNA double-strand break repair Mre11 nuclease and Rad50-ATPase. Cell 420 11371344
1997 hMre11 and hRad50 nuclear foci are induced during the normal cellular response to DNA double-strand breaks. Molecular and cellular biology 413 9315668
2010 DNA end resection by Dna2-Sgs1-RPA and its stimulation by Top3-Rmi1 and Mre11-Rad50-Xrs2. Nature 383 20811461
2007 Mre11-Rad50-Nbs1 is a keystone complex connecting DNA repair machinery, double-strand break signaling, and the chromatin template. Biochemistry and cell biology = Biochimie et biologie cellulaire 319 17713585
2018 The MRE11-RAD50-NBS1 Complex Conducts the Orchestration of Damage Signaling and Outcomes to Stress in DNA Replication and Repair. Annual review of biochemistry 318 29709199
2009 Nbs1 flexibly tethers Ctp1 and Mre11-Rad50 to coordinate DNA double-strand break processing and repair. Cell 265 19804755
1993 DNA double-strand breaks and the RAD50-RAD57 genes in Saccharomyces. Seminars in cancer biology 232 8513150
2001 DNA structure-specific nuclease activities in the Saccharomyces cerevisiae Rad50*Mre11 complex. The Journal of biological chemistry 225 11454871
2005 Mesoscale conformational changes in the DNA-repair complex Rad50/Mre11/Nbs1 upon binding DNA. Nature 215 16163361
2019 MRE11-RAD50-NBS1 complex alterations and DNA damage response: implications for cancer treatment. Molecular cancer 205 31767017
2010 Mre11-Rad50-Xrs2 and Sae2 promote 5' strand resection of DNA double-strand breaks. Nature structural & molecular biology 190 21102445
2006 Two-step activation of ATM by DNA and the Mre11-Rad50-Nbs1 complex. Nature structural & molecular biology 187 16622404
1996 Human Rad50 is physically associated with human Mre11: identification of a conserved multiprotein complex implicated in recombinational DNA repair. Molecular and cellular biology 185 8756642
2010 Saccharomyces cerevisiae Mre11/Rad50/Xrs2 and Ku proteins regulate association of Exo1 and Dna2 with DNA breaks. The EMBO journal 181 20834227
2010 Mre11-Rad50-Nbs1 conformations and the control of sensing, signaling, and effector responses at DNA double-strand breaks. DNA repair 180 21035407
2003 Rad50/SMC proteins and ABC transporters: unifying concepts from high-resolution structures. Current opinion in structural biology 174 12727520
2002 Cancer predisposition and hematopoietic failure in Rad50(S/S) mice. Genes & development 168 12208847
2006 RAD50 and NBS1 are breast cancer susceptibility genes associated with genomic instability. Carcinogenesis 159 16474176
2017 Single-Molecule Imaging Reveals How Mre11-Rad50-Nbs1 Initiates DNA Break Repair. Molecular cell 152 28867292
2016 Nbs1 Converts the Human Mre11/Rad50 Nuclease Complex into an Endo/Exonuclease Machine Specific for Protein-DNA Adducts. Molecular cell 141 27814491
2005 The Mre11/Rad50/Nbs1 complex and its role as a DNA double-strand break sensor for ATM. Cell cycle (Georgetown, Tex.) 131 15908798
2014 ATP-driven Rad50 conformations regulate DNA tethering, end resection, and ATM checkpoint signaling. The EMBO journal 130 24493214
2014 Rad50-CARD9 interactions link cytosolic DNA sensing to IL-1β production. Nature immunology 125 24777530
2004 Replication protein A and the Mre11.Rad50.Nbs1 complex co-localize and interact at sites of stalled replication forks. The Journal of biological chemistry 125 15180989
2001 The forkhead-associated domain of NBS1 is essential for nuclear foci formation after irradiation but not essential for hRAD50[middle dot]hMRE11[middle dot]NBS1 complex DNA repair activity. The Journal of biological chemistry 120 11062235
2011 Crystal structure of the Mre11-Rad50-ATPγS complex: understanding the interplay between Mre11 and Rad50. Genes & development 119 21511873
2002 Tethering on the brink: the evolutionarily conserved Mre11-Rad50 complex. Trends in biochemical sciences 113 12151226
2018 Physiological protein blocks direct the Mre11-Rad50-Xrs2 and Sae2 nuclease complex to initiate DNA end resection. Genes & development 110 29321179
2004 MRE11/RAD50/NBS1: complex activities. Chromosoma 109 15309560
2019 Mechanism of DNA End Sensing and Processing by the Mre11-Rad50 Complex. Molecular cell 107 31492634
2000 Mre11 and Rad50 from Pyrococcus furiosus: cloning and biochemical characterization reveal an evolutionarily conserved multiprotein machine. Journal of bacteriology 106 11029422
2018 Plasticity of the Mre11-Rad50-Xrs2-Sae2 nuclease ensemble in the processing of DNA-bound obstacles. Genes & development 103 29321177
2015 ATP-dependent DNA binding, unwinding, and resection by the Mre11/Rad50 complex. The EMBO journal 100 26717941
2001 Fission yeast Rad50 stimulates sister chromatid recombination and links cohesion with repair. The EMBO journal 98 11726502
2004 The Drosophila Mre11/Rad50 complex is required to prevent both telomeric fusion and chromosome breakage. Current biology : CB 97 15296753
2007 MRE11-RAD50-NBS1 and ATM function as co-mediators of TRF1 in telomere length control. Nature structural & molecular biology 94 17694070
2015 Envisioning the dynamics and flexibility of Mre11-Rad50-Nbs1 complex to decipher its roles in DNA replication and repair. Progress in biophysics and molecular biology 89 25576492
2007 Rad50 adenylate kinase activity regulates DNA tethering by Mre11/Rad50 complexes. Molecular cell 87 17349953
2004 A bipolar DNA helicase gene, herA, clusters with rad50, mre11 and nurA genes in thermophilic archaea. Nucleic acids research 87 14990749
2016 Structural mechanism of ATP-dependent DNA binding and DNA end bridging by eukaryotic Rad50. The EMBO journal 86 26896444
2003 Yeast xrs2 binds DNA and helps target rad50 and mre11 to DNA ends. The Journal of biological chemistry 86 14522986
2009 MRE11-RAD50-NBS1 complex dictates DNA repair independent of H2AX. The Journal of biological chemistry 84 19910469
2002 DNA cross-link-dependent RAD50/MRE11/NBS1 subnuclear assembly requires the Fanconi anemia C protein. Human molecular genetics 80 12354779
2019 MRE11-RAD50-NBS1 promotes Fanconi Anemia R-loop suppression at transcription-replication conflicts. Nature communications 75 31537797
2004 The rad50 signature motif: essential to ATP binding and biological function. Journal of molecular biology 75 14698290
2004 Molecular mechanism of the recruitment of NBS1/hMRE11/hRAD50 complex to DNA double-strand breaks: NBS1 binds to gamma-H2AX through FHA/BRCT domain. Journal of radiation research 75 15635255
2001 Cdc13 prevents telomere uncapping and Rad50-dependent homologous recombination. The EMBO journal 73 11689452
2011 ATM protein-dependent phosphorylation of Rad50 protein regulates DNA repair and cell cycle control. The Journal of biological chemistry 71 21757780
2010 Making the best of the loose ends: Mre11/Rad50 complexes and Sae2 promote DNA double-strand break resection. DNA repair 71 21050828
2016 Xrs2 Dependent and Independent Functions of the Mre11-Rad50 Complex. Molecular cell 66 27746018
2019 C1QBP Promotes Homologous Recombination by Stabilizing MRE11 and Controlling the Assembly and Activation of MRE11/RAD50/NBS1 Complex. Molecular cell 64 31353207
2005 MRE11/RAD50 cleaves DNA in the AID/UNG-dependent pathway of immunoglobulin gene diversification. Molecular cell 64 16285919
2001 Frameshift mutations at coding mononucleotide repeats of the hRAD50 gene in gastrointestinal carcinomas with microsatellite instability. Cancer research 64 11196187
2017 Eukaryotic Rad50 functions as a rod-shaped dimer. Nature structural & molecular biology 63 28134932
2004 Independent roles for nibrin and Mre11-Rad50 in the activation and function of Atm. The Journal of biological chemistry 63 15234984
2014 Structure of the Rad50 DNA double-strand break repair protein in complex with DNA. The EMBO journal 59 25349191
2013 DNA-dependent protein kinase regulates DNA end resection in concert with Mre11-Rad50-Nbs1 (MRN) and ataxia telangiectasia-mutated (ATM). The Journal of biological chemistry 59 24220101
2009 RAD50, an SMC family member with multiple roles in DNA break repair: how does ATP affect function? Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology 57 19308707
2019 Stepwise 5' DNA end-specific resection of DNA breaks by the Mre11-Rad50-Xrs2 and Sae2 nuclease ensemble. Proceedings of the National Academy of Sciences of the United States of America 56 30819891
2013 Visualization of local DNA unwinding by Mre11/Rad50/Nbs1 using single-molecule FRET. Proceedings of the National Academy of Sciences of the United States of America 56 24191051
2000 RINT-1, a novel Rad50-interacting protein, participates in radiation-induced G(2)/M checkpoint control. The Journal of biological chemistry 56 11096100
2007 Structural conservation of RecF and Rad50: implications for DNA recognition and RecF function. The EMBO journal 54 17255941
2009 MRE11-RAD50-NBS1 is a critical regulator of FANCD2 stability and function during DNA double-strand break repair. The EMBO journal 52 19609304
2008 Dancing on damaged chromatin: functions of ATM and the RAD50/MRE11/NBS1 complex in cellular responses to DNA damage. Journal of radiation research 51 18772547
2005 The MRE11-RAD50-NBS1 complex accelerates somatic hypermutation and gene conversion of immunoglobulin variable regions. Nature immunology 51 15937485
2002 DNA end-binding specificity of human Rad50/Mre11 is influenced by ATP. Nucleic acids research 51 12384589
2022 Cryo-EM structure of the Mre11-Rad50-Nbs1 complex reveals the molecular mechanism of scaffolding functions. Molecular cell 50 36577401
2010 Biochemical characterization of bacteriophage T4 Mre11-Rad50 complex. The Journal of biological chemistry 50 21081488
2005 Rad50 depletion impacts upon ATR-dependent DNA damage responses. Human molecular genetics 48 16087684
2009 RAD50 and NBS1 form a stable complex functional in DNA binding and tethering. Nucleic acids research 47 19151086
2005 The Mre11/Rad50/Xrs2 complex and non-homologous end-joining of incompatible ends in S. cerevisiae. DNA repair 45 16043424
2014 The Rad50 hook domain regulates DNA damage signaling and tumorigenesis. Genes & development 44 24532689
2015 Interdependence of the rad50 hook and globular domain functions. Molecular cell 43 25601756
2014 The Mre11/Rad50/Nbs1 complex: recent insights into catalytic activities and ATP-driven conformational changes. Experimental cell research 40 25016281
2008 Rad50 is dispensable for the maintenance and viability of postmitotic tissues. Molecular and cellular biology 39 19001091
2002 The plant Rad50-Mre11 protein complex. FEBS letters 39 11959125
2008 ATM and the Mre11-Rad50-Nbs1 complex respond to nucleoside analogue-induced stalled replication forks and contribute to drug resistance. Cancer research 38 18829552
2022 Mre11-Rad50 oligomerization promotes DNA double-strand break repair. Nature communications 36 35501303
2019 The ATP-bound conformation of the Mre11-Rad50 complex is essential for Tel1/ATM activation. Nucleic acids research 35 30698745
2011 MRE11 and RAD50, but not NBS1, are essential for gene targeting in the moss Physcomitrella patens. Nucleic acids research 35 22210882
2006 Purification and biochemical characterization of ataxia-telangiectasia mutated and Mre11/Rad50/Nbs1. Methods in enzymology 34 16793391
2022 Structural mechanism of endonucleolytic processing of blocked DNA ends and hairpins by Mre11-Rad50. Molecular cell 33 35987200
2017 RAD50 Expression Is Associated with Poor Clinical Outcomes after Radiotherapy for Resected Non-small Cell Lung Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research 33 29030353
2011 ATP hydrolysis by RAD50 protein switches MRE11 enzyme from endonuclease to exonuclease. The Journal of biological chemistry 33 22102415
2019 Activation of Tel1ATM kinase requires Rad50 ATPase and long nucleosome-free DNA but no DNA ends. The Journal of biological chemistry 31 31073030
2017 Rad50 ATPase activity is regulated by DNA ends and requires coordination of both active sites. Nucleic acids research 31 28369545
2012 Ataxia-telangiectasia mutated and the Mre11-Rad50-NBS1 complex: promising targets for radiosensitization. Acta medica Okayama 31 22525466
2020 A Survey of Reported Disease-Related Mutations in the MRE11-RAD50-NBS1 Complex. Cells 30 32668560
2012 Adenovirus regulates sumoylation of Mre11-Rad50-Nbs1 components through a paralog-specific mechanism. Journal of virology 30 22740413
2023 RBBP4 regulates the expression of the Mre11-Rad50-NBS1 (MRN) complex and promotes DNA double-strand break repair to mediate glioblastoma chemoradiotherapy resistance. Cancer letters 28 36736531
2020 Human RAD50 deficiency: Confirmation of a distinctive phenotype. American journal of medical genetics. Part A 28 32212377
2006 Two roles for Rad50 in telomere maintenance. The EMBO journal 28 16990794
2003 Differential involvement of the hMRE11/hRAD50/NBS1 complex, BRCA1 and MLH1 in NF-kappaB activation by camptothecin and X-ray. Oncogene 27 12955088
2014 RAD50 phosphorylation promotes ATR downstream signaling and DNA restart following replication stress. Human molecular genetics 26 24694934

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