Interaction of hREV1 with three human Y-family DNA polymerases.
Polkappa is one of many DNA polymerases involved in translesion DNA synthesis (TLS). It belongs to the Y-family of polymerases along with Poleta, Poliota and hREV1. Unlike Poleta encoded by the xeroderma pigmentosum variant (XPV) gene, Polkappa is unable to bypass UV-induced DNA damage in vitro, but it is able to bypass benzo[a]pyrene (B[a]P)-adducted guanines accurately and efficiently. In an attempt to identify factor(s) targeting Polkappa to its cognate DNA lesion(s), we searched for Polkappa-interacting proteins by using the yeast two-hybrid assay. We found that Polkappa interacts with a C-terminal region of hREV1. Poleta and Poliota were also found to interact with the same region of hREV1. The interaction between Polkappa and hREV1 was confirmed by pull-down and co-immunoprecipitation assays. The C-terminal region of hREV1 is known to interact with hREV7, a non-catalytic subunit of Polzeta that is another structurally unrelated TLS enzyme, and we show that Polkappa and hREV7 bind to the same C-terminal region of hREV1. Thus, our results suggest that hREV1 plays a pivotal role in the multi-enzyme, multi-step process of translesion DNA synthesis.
Ohashi
Eiji
E
Institute for Virus Research, Kyoto University, 53 Shogoin-Kawaracho, Sakyo-ku, Kyoto 606-8507, Japan.
Murakumo
Yoshiki
Y
Kanjo
Naoko
N
Akagi
Jun-Ichi
J
Masutani
Chikahide
C
Hanaoka
Fumio
F
Ohmori
Haruo
H
D000595:Amino Acid Sequence; D001564:Benzo(a)pyrene / Q000494:pharmacology; D002460:Cell Line; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006147:Guanine / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D020798:Two-Hybrid System Techniques; D014466:Ultraviolet Rays; D001616:beta-Galactosidase / Q000378:metabolism; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
D001564:Benzo(a)pyrene; D004259:DNA-Directed DNA Polymerase; D005982:Glutathione Transferase; D006147:Guanine; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D000094769:DNA Polymerase iota; C429004:REV1 protein, human; D000099270:Y-Family DNA Polymerases; C404302:POLK protein, human