Rev1 is a base excision repair enzyme with 5'-deoxyribose phosphate lyase activity.
Rev1 is a member of the Y-family of DNA polymerases and is known for its deoxycytidyl transferase activity that incorporates dCMP into DNA and its ability to function as a scaffold factor for other Y-family polymerases in translesion bypass events. Rev1 also is involved in mutagenic processes during somatic hypermutation of immunoglobulin genes. In light of the mutation pattern consistent with dCMP insertion observed earlier in mouse fibroblast cells treated with a base excision repair-inducing agent, we questioned whether Rev1 could also be involved in base excision repair (BER). Here, we uncovered a weak 5'-deoxyribose phosphate (5'-dRP) lyase activity in mouse Rev1 and demonstrated the enzyme can mediate BER in vitro The full-length Rev1 protein and its catalytic core domain are similar in their ability to support BER in vitro The dRP lyase activity in both of these proteins was confirmed by NaBH4reduction of the Schiff base intermediate and kinetics studies. Limited proteolysis, mass spectrometry and deletion analysis localized the dRP lyase active site to the C-terminal segment of Rev1's catalytic core domain. These results suggest that Rev1 could serve as a backup polymerase in BER and could potentially contribute to AID-initiated antibody diversification through this activity.
Genome Integrity and Structural Biology Laboratory, National Institutes of Health, NIEHS, 111 T.W. Alexander Drive, PO Box 12233, MD F3-01, Research Triangle Park, NC 27709, USA.
Rajendra
R
Prasad
Genome Integrity and Structural Biology Laboratory, National Institutes of Health, NIEHS, 111 T.W. Alexander Drive, PO Box 12233, MD F3-01, Research Triangle Park, NC 27709, USA.
Vladimir
V
Poltoratsky
Genome Integrity and Structural Biology Laboratory, National Institutes of Health, NIEHS, 111 T.W. Alexander Drive, PO Box 12233, MD F3-01, Research Triangle Park, NC 27709, USA.
Esther W
EW
Hou
Genome Integrity and Structural Biology Laboratory, National Institutes of Health, NIEHS, 111 T.W. Alexander Drive, PO Box 12233, MD F3-01, Research Triangle Park, NC 27709, USA wilson5@niehs.nih.gov.
Samuel H
SH
Wilson
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D004260:DNA Repair; D004259:DNA-Directed DNA Polymerase; D007700:Kinetics; D051379:Mice; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000502:physiology*; D010449:Peptide Mapping; D019761:Phosphorus-Oxygen Lyases / Q000737:chemistry / Q000502:physiology; D059748:Proteolysis; D000099270:Y-Family DNA Polymerases
D004259:DNA-Directed DNA Polymerase; D009713:Nucleotidyltransferases; D019761:Phosphorus-Oxygen Lyases; C423906:5'-deoxyribose phosphate lyase; C496086:Rev1 protein, mouse; D000099270:Y-Family DNA Polymerases
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