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PMID: 15520275 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Idling by DNA polymerase delta maintains a ligatable nick during lagging-strand DNA replication.

Genes & development ·Vol. 18 ·No. 22 ·2004-11-15 ·Pages 2764-73

Garg P, Stith CM, Sabouri N, Johansson E, Burgers PM

Abstract

During each yeast cell cycle, approximately 100,000 nicks are generated during lagging-strand DNA replication. Efficient nick processing during Okazaki fragment maturation requires the coordinated action of DNA polymerase delta (Pol delta) and the FLAP endonuclease FEN1. Misregulation of this process leads to the accumulation of double-stranded breaks and cell lethality. Our studies highlight a remarkably efficient mechanism for Okazaki fragment maturation in which Pol delta by default displaces 2-3 nt of any downstream RNA or DNA it encounters. In the presence of FEN1, efficient nick translation ensues, whereby a mixture of mono- and small oligonucleotides are released. If FEN1 is absent or not optimally functional, the ability of Pol delta to back up via its 3'-5'-exonuclease activity, a process called idling, maintains the polymerase at a position that is ideal either for ligation (in case of a DNA-DNA nick) or for subsequent engagement by FEN1 (in case of a DNA-RNA nick). Consistent with the hypothesis that DNA polymerase epsilon is the leading-strand enzyme, we observed no idling by this enzyme and no cooperation with FEN1 for creating a ligatable nick.

MeSH Terms
DNA/metabolism DNA Helicases/metabolism DNA Polymerase III/metabolism DNA Primers DNA Replication Exonucleases/metabolism Flap Endonucleases/metabolism Oligonucleotides/metabolism Saccharomyces cerevisiae/enzymology,genetics
Chemicals
DNA Primers Oligonucleotides DNA DNA Polymerase III Exonucleases Flap Endonucleases DNA Helicases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Garg Parie
Department of Biochemistry, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Stith Carrie M
Sabouri Nasim
Johansson Erik
Burgers Peter M
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1252304
Published
2004-11-15
Epub
2004-00-01
Pages
2764-73
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC528896
Subset
IM
Grants
NIGMS NIH HHS · R01 GM032431 · United States
NIGMS NIH HHS · GM32431 · United States
External Links
PubMed source DOI: 10.1101/gad.1252304 PMC Full Text View journal details
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