title: The Rev1 translesion synthesis polymerase has multiple distinct DNA binding modes.
abstract: Rev1 is a eukaryotic DNA polymerase of the Y family involved in translesion synthesis (TLS), a major damage tolerance pathway that allows DNA replication at damaged templates. Uniquely amongst the Y family polymerases, the N-terminal part of Rev1, dubbed the BRCA1 C-terminal homology (BRCT) region, includes a BRCT domain. While most BRCT domains mediate protein-protein interactions, Rev1 contains a predicted α-helix N-terminal to the BRCT domain and in human Replication Factor C (RFC) such a BRCT region endows the protein with DNA binding capacity. Here, we studied the DNA binding properties of yeast and mouse Rev1. Our results show that the BRCT region of Rev1 specifically binds to a 5' phosphorylated, recessed, primer-template junction. This DNA binding depends on the extra α-helix, N-terminal to the BRCT domain. Surprisingly, a stretch of 20 amino acids N-terminal to the predicted α-helix is also critical for high-affinity DNA binding. In addition to 5' primer-template junction binding, Rev1 efficiently binds to a recessed 3' primer-template junction. These dual DNA binding characteristics are discussed in view of the proposed recruitment of Rev1 by 5' primer-template junctions, downstream of stalled replication forks.
authors.affiliation: Department of Protein Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratory, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
authors.forename: Frederik H
authors.initials: FH
authors.lastname: de Groote
authors.forename: Jacob G
authors.initials: JG
authors.lastname: Jansen
authors.forename: Yuji
authors.initials: Y
authors.lastname: Masuda
authors.forename: Dipen M
authors.initials: DM
authors.lastname: Shah
authors.forename: Kenji
authors.initials: K
authors.lastname: Kamiya
authors.forename: Niels
authors.initials: N
authors.lastname: de Wind
authors.forename: Gregg
authors.initials: G
authors.lastname: Siegal
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D000099270:Y-Family DNA Polymerases
chemical_list: D004247:DNA; D004259:DNA-Directed DNA Polymerase; D009713:Nucleotidyltransferases; D029701:Saccharomyces cerevisiae Proteins; C100951:REV1 protein, S cerevisiae; D000099270:Y-Family DNA Polymerases; C496086:Rev1 protein, mouse