Vertebrate DNA damage tolerance requires the C-terminus but not BRCT or transferase domains of REV1.
REV1 is central to the DNA damage response of eukaryotes through an as yet poorly understood role in translesion synthesis. REV1 is a member of the Y-type DNA polymerase family and is capable of in vitro deoxycytidyl transferase activity opposite a range of damaged bases. However, non-catalytic roles for REV1 have been suggested by the Saccharomyces cerevisiae rev1-1 mutant, which carries a point mutation in the N-terminal BRCT domain, and the recently demonstrated ability of the mammalian protein to interact with each of the other translesion polymerases via its extreme C-terminus. Here, we show that a region adjacent to this polymerase interacting domain mediates an interaction with PCNA. These C-terminal domains of REV1 are necessary, although not sufficient, for effective tolerance of DNA damage in the avian cell line DT40, while the BRCT domain and transferase activity are not directly required. Together these data provide strong support for REV1 playing an important non-catalytic role in coordinating translesion synthesis. Further, unlike in budding yeast, rad18 is not epistatic to rev1 for DNA damage tolerance suggesting that REV1 and RAD18 play largely independent roles in the control of vertebrate translesion synthesis.
Ross
Anna-Laura
AL
Medical Research Council Laboratory of Molecular Biology Hills Road, Cambridge, CB2 2QH, UK.
Simpson
Laura J
LJ
Sale
Julian E
JE
D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry; D002645:Chickens / Q000235:genetics; D002945:Cisplatin / Q000633:toxicity; D017124:Conserved Sequence; D004247:DNA / Q000096:biosynthesis; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000502:physiology; D005816:Genetic Complementation Test; D009154:Mutation; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D017434:Protein Structure, Tertiary; D014466:Ultraviolet Rays; D014714:Vertebrates / Q000235:genetics*; D000099270:Y-Family DNA Polymerases
D002945:Cisplatin; D004247:DNA; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D009713:Nucleotidyltransferases; D018809:Proliferating Cell Nuclear Antigen; C429004:REV1 protein, human; D000099270:Y-Family DNA Polymerases
Nucleic Acids Res. 1999 Nov 15;27(22):4468-75
10536157
Genetics. 1997 Dec;147(4):1557-68
9409821
Genes Dev. 2001 Apr 15;15(8):945-54
11316789
Mol Cell. 2004 May 21;14(4):491-500
15149598
BMC Biotechnol. 2001;1:7
11591226
Philos Trans R Soc Lond B Biol Sci. 2001 Jan 29;356(1405):41-6
11205328
EMBO J. 2003 Dec 15;22(24):6621-30
14657033
EMBO J. 2002 Oct 15;21(20):5558-66
12374756
J Biol Chem. 2001 May 4;276(18):15051-8
11278384
Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4186-91
10760286
Nature. 2003 Sep 11;425(6954):188-91
12968183
Nucleic Acids Res. 2002 Apr 1;30(7):1630-8
11917024
Nature. 2003 Oct 16;425(6959):737-41
14562106
Nucleic Acids Res. 2002 Dec 1;30(23):5129-35
12466536
J Biol Chem. 2001 Sep 21;276(38):35644-51
11485998
Genetics. 1976 Feb;82(2):207-32
770231
Genes Cells. 2004 Jun;9(6):523-31
15189446
Mol Cell Biol. 2001 Feb;21(3):713-20
11154259
Science. 2003 Oct 24;302(5645):636-9
14576432
DNA Repair (Amst). 2003 Oct 7;2(10):1075-85
13679146
EMBO J. 2003 Jun 16;22(12):3188-97
12805232
J Biol Chem. 2002 Dec 13;277(50):48690-5
12356753
Mol Cell Biol. 2003 Jan;23(2):754-61
12509472
Bioessays. 2000 Nov;22(11):997-1006
11056476
Nucleic Acids Res Suppl. 2002;(2):87-8
12903118
Annu Rev Biochem. 2002;71:17-50
12045089
EMBO J. 2004 Oct 1;23(19):3886-96
15359278
DNA Repair (Amst). 2002 Jun 21;1(6):425-35
12509231
Nature. 2002 Sep 12;419(6903):135-41
12226657
J Mol Biol. 2000 Jun 2;299(2):499-520
10860755
Nucleic Acids Res. 2005;33(1):356-65
15653636
J Mol Biol. 1995 Aug 11;251(2):229-36
7643399
FEBS Lett. 1997 Jan 2;400(1):25-30
9000507
Biochem J. 2004 Sep 1;382(Pt 2):535-43
15175013
Mol Cell Biol. 2003 Jan;23(2):474-81
12509447
FEBS Lett. 2002 Jun 5;520(1-3):88-92
12044876
Mol Cell Biol. 2004 May;24(10):4267-74
15121847
Nature. 1996 Aug 22;382(6593):729-31
8751446
Mol Microbiol. 2000 Aug;37(3):549-54
10931348
DNA Repair (Amst). 2004 Nov 2;3(11):1503-14
15380106
Genetics. 1971 May;68(1):21-33
17248528
J Bacteriol. 1989 Jan;171(1):230-7
2492497
Science. 2003 Oct 24;302(5645):639-42
14576433
EMBO J. 2003 Apr 1;22(7):1654-64
12660171