Domain structure, localization, and function of DNA polymerase eta, defective in xeroderma pigmentosum variant cells.
DNA polymerase eta carries out translesion synthesis past UV photoproducts and is deficient in xeroderma pigmentosum (XP) variants. We report that poleta is mostly localized uniformly in the nucleus but is associated with replication foci during S phase. Following treatment of cells with UV irradiation or carcinogens, it accumulates at replication foci stalled at DNA damage. The C-terminal third of poleta is not required for polymerase activity. However, the C-terminal 70 aa are needed for nuclear localization and a further 50 aa for relocalization into foci. Poleta truncations lacking these domains fail to correct the defects in XP-variant cells. Furthermore, we have identified mutations in two XP variant patients that leave the polymerase motifs intact but cause loss of the localization domains.
Kannouche
P
P
MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton BN1 9RR, UK.
Broughton
B C
BC
Volker
M
M
Hanaoka
F
F
Mullenders
L H
LH
Lehmann
A R
AR
D000595:Amino Acid Sequence; D001483:Base Sequence; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000201:enzymology; D004247:DNA / Q000378:metabolism / Q000528:radiation effects; D004249:DNA Damage; D017931:DNA Primers / Q000235:genetics; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014644:Genetic Variation; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051135:Rad51 Recombinase; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014466:Ultraviolet Rays / Q000009:adverse effects; D014983:Xeroderma Pigmentosum / Q000201:enzymology* / Q000235:genetics*; D000099270:Y-Family DNA Polymerases
D004247:DNA; D017931:DNA Primers; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D018809:Proliferating Cell Nuclear Antigen; D051135:Rad51 Recombinase; D011993:Recombinant Fusion Proteins; C495153:RAD51 protein, human; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases