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TitleIdentification of fidelity-governing factors in human recombinases DMC1 and RAD51 from cryo-EM structures.
Journal, issue, pagesNat Commun, Vol. 12, Issue 1, Page 115, Year 2021
Publish dateJan 14, 2021
AuthorsShih-Chi Luo / Hsin-Yi Yeh / Wei-Hsuan Lan / Yi-Min Wu / Cheng-Han Yang / Hao-Yen Chang / Guan-Chin Su / Chia-Yi Lee / Wen-Jin Wu / Hung-Wen Li / Meng-Chiao Ho / Peter Chi / Ming-Daw Tsai /
PubMed AbstractBoth high-fidelity and mismatch-tolerant recombination, catalyzed by RAD51 and DMC1 recombinases, respectively, are indispensable for genomic integrity. Here, we use cryo-EM, MD simulation and ...Both high-fidelity and mismatch-tolerant recombination, catalyzed by RAD51 and DMC1 recombinases, respectively, are indispensable for genomic integrity. Here, we use cryo-EM, MD simulation and functional analysis to elucidate the structural basis for the mismatch tolerance of DMC1. Structural analysis of DMC1 presynaptic and postsynaptic complexes suggested that the lineage-specific Loop 1 Gln244 (Met243 in RAD51) may help stabilize DNA backbone, whereas Loop 2 Pro274 and Gly275 (Val273/Asp274 in RAD51) may provide an open "triplet gate" for mismatch tolerance. In support, DMC1-Q244M displayed marked increase in DNA dynamics, leading to unobservable DNA map. MD simulation showed highly dispersive mismatched DNA ensemble in RAD51 but well-converged DNA in DMC1 and RAD51-V273P/D274G. Replacing Loop 1 or Loop 2 residues in DMC1 with RAD51 counterparts enhanced DMC1 fidelity, while reciprocal mutations in RAD51 attenuated its fidelity. Our results show that three Loop 1/Loop 2 residues jointly enact contrasting fidelities of DNA recombinases.
External linksNat Commun / PubMed:33446654 / PubMed Central
MethodsEM (helical sym.)
Resolution3.2 - 3.47 Å
Structure data

30308

7c98

EMDB-30308, PDB-7c98:
Human DMC1 post-synaptic complexes
Method: EM (helical sym.) / Resolution: 3.47 Å

30309

7c99

EMDB-30309, PDB-7c99:
Human DMC1 post-synaptic complexes with mismatched dsDNA
Method: EM (helical sym.) / Resolution: 3.36 Å

30310

7c9a

EMDB-30310, PDB-7c9a:
Human RAD51 post-synaptic complexes mutant (V273P, D274G)
Method: EM (helical sym.) / Resolution: 3.43 Å

30311

7c9c

EMDB-30311, PDB-7c9c:
Human DMC1 pre-synaptic complexes
Method: EM (helical sym.) / Resolution: 3.33 Å

30366

7cgy

EMDB-30366, PDB-7cgy:
Human DMC1 Q244M mutant of the post-synaptic complexes
Method: EM (helical sym.) / Resolution: 3.2 Å

Chemicals

ChemComp-CA:
Unknown entry

ChemComp-ANP:
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMP-PNP, energy-carrying molecule analogue*YM

Source
  • homo sapiens (human)
  • synthetic construct (others)
KeywordsRECOMBINATION/DNA / meiotic homologous recombination / DNA repair / ATPase / RECOMBINATION / RECOMBINATION-DNA complex / meitotic homologous recombination

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