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PMID: 12426396 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Localization of DNA polymerases eta and iota to the replication machinery is tightly co-ordinated in human cells.

The EMBO journal ·Vol. 21 ·No. 22 ·2002-11-15 ·Pages 6246-56

Kannouche P, Fernández de Henestrosa AR, Coull B, Vidal AE, Gray C, Zicha D, Woodgate R, Lehmann AR

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.

MeSH Terms
Active Transport, Cell Nucleus Animals Caffeine/toxicity Cell Line Cell Line, Transformed/drug effects,radiation effects Cell Nucleus/enzymology DNA/drug effects,radiation effects DNA Damage DNA Replication/physiology DNA-Directed DNA Polymerase/chemistry,deficiency,genetics,physiology Genes, Reporter Genetic Complementation Test Humans Microscopy, Fluorescence Protein Interaction Mapping Recombinant Fusion Proteins/metabolism Sequence Deletion Spodoptera/cytology Transfection Two-Hybrid System Techniques Ultraviolet Rays Xeroderma Pigmentosum/enzymology,genetics,pathology
Chemicals
Recombinant Fusion Proteins Caffeine DNA DNA polymerase iota DNA-Directed DNA Polymerase Rad30 protein
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Kannouche Patricia
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.
Fernández de Henestrosa Antonio R
Coull Barry
Vidal Antonio E
Gray Colin
Zicha Daniel
Woodgate Roger
Lehmann Alan R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
2002-11-15
Pages
6246-56
Language
English
Region
England
NLM ID
8208664
PMCID
PMC137208
Subset
IM
Corrections
RepublishedIn
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