title: Localization of DNA polymerases eta and iota to the replication machinery is tightly co-ordinated in human cells.
abstract: Y-family DNA polymerases can replicate past a variety of damaged bases in vitro but, with the exception of DNA polymerase eta (poleta), which is defective in xeroderma pigmentosum variants, there is little information on the functions of these polymerases in vivo. Here, we show that DNA polymerase iota (poliota), like poleta, associates with the replication machinery and accumulates at stalled replication forks following DNA-damaging treatment. We show that poleta and poliota foci form with identical kinetics and spatial distributions, suggesting that localization of these two polymerases is tightly co-ordinated within the nucleus. Furthermore, localization of poliota in replication foci is largely dependent on the presence of poleta. Using several different approaches, we demonstrate that poleta and poliota interact with each other physically and that the C-terminal 224 amino acids of poliota are sufficient for both the interaction with poleta and accumulation in replication foci. Our results provide strong evidence that poleta targets poliota to the replication machinery, where it may play a general role in maintaining genome integrity as well as participating in translesion DNA synthesis.
authors.affiliation: Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton BN1 9RQ, Cancer Research UK London Research Institute, 44, Lincoln's Inn Fields, London WC2A 3PX, UK.
authors.forename: Patricia
authors.initials: P
authors.lastname: Kannouche
authors.forename: Antonio R
authors.initials: AR
authors.lastname: Fernández de Henestrosa
authors.forename: Barry
authors.initials: B
authors.lastname: Coull
authors.forename: Antonio E
authors.initials: AE
authors.lastname: Vidal
authors.forename: Colin
authors.initials: C
authors.lastname: Gray
authors.forename: Daniel
authors.initials: D
authors.lastname: Zicha
authors.forename: Roger
authors.initials: R
authors.lastname: Woodgate
authors.forename: Alan R
authors.initials: AR
authors.lastname: Lehmann
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002110:Caffeine / Q000633:toxicity; D002460:Cell Line; D002461:Cell Line, Transformed / Q000187:drug effects / Q000528:radiation effects; D002467:Cell Nucleus / Q000201:enzymology; D004247:DNA / Q000187:drug effects / Q000528:radiation effects; D004249:DNA Damage; D004261:DNA Replication / Q000502:physiology*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D006801:Humans; D008856:Microscopy, Fluorescence; D025941:Protein Interaction Mapping; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D018411:Spodoptera / Q000166:cytology; D014162:Transfection; D020798:Two-Hybrid System Techniques; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
chemical_list: D002110:Caffeine; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D011993:Recombinant Fusion Proteins; D000094769:DNA Polymerase iota; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases