Localisation of human Y-family DNA polymerase kappa: relationship to PCNA foci.
DNA polymerases of the Y-family are involved in translesion DNA synthesis past different types of DNA damage. Previous work has shown that DNA polymerases eta and iota are localised in replication factories during S phase, where they colocalise one-to-one with PCNA. Cells with factories containing these polymerases accumulate after treatment with DNA damaging agents because replication forks are stalled at sites of damage. We now show that DNA polymerase kappa (pol(kappa)) has a different localisation pattern. Although, like the other Y-family polymerases, it is exclusively localised in the nucleus, pol(kappa) is found in replication foci in only a small proportion of S-phase cells. It does not colocalise in those foci with proliferating cell nuclear antigen (PCNA) in the majority of cells. This reduced number of cells with pol(kappa) foci, when compared with those containing pol(eta) foci, is observed both in untreated cells and in cells treated with hydroxyurea, UV irradiation or benzo[a]pyrene. The C-terminal 97 amino acids of pol(kappa)are sufficient for this limited localisation into nuclear foci, and include a C2HC zinc finger, bipartite nuclear localisation signal and putative PCNA binding site.
Ogi
Tomoo
T
Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton, BN1 9RR, UK.
Kannouche
Patricia
P
Lehmann
Alan R
AR
D000595:Amino Acid Sequence; D001564:Benzo(a)pyrene / Q000494:pharmacology; D015153:Blotting, Western; D002273:Carcinogens / Q000494:pharmacology; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002472:Cell Transformation, Viral; D004249:DNA Damage / Q000187:drug effects / Q000528:radiation effects; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000528:radiation effects; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D013347:Subcellular Fractions; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
D001564:Benzo(a)pyrene; D002273:Carcinogens; D004259:DNA-Directed DNA Polymerase; D049452:Green Fluorescent Proteins; D006918:Hydroxyurea; D019384:Nucleic Acid Synthesis Inhibitors; D018809:Proliferating Cell Nuclear Antigen; C404302:POLK protein, human; D000099270:Y-Family DNA Polymerases