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Idling by DNA polymerase delta maintains a ligatable nick during lagging-strand DNA replication.
- Garg, Parie (author)
- Stith, Carrie M (author)
-
- Sabouri, Nasim (author)
- Umeå universitet,Institutionen för medicinsk kemi och biofysik
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- Burgers, Peter M (author)
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- (creator_code:org_t)
- 2004-11-01
- 2004
- English.
- In: Genes & Development. - : Cold Spring Harbor Laboratory. - 0890-9369 .- 1549-5477. ; 18:22, s. 2764-2773
- Journal article (peer-reviewed)
Abstract
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- During each yeast cell cycle, ∼100,000 nicks are generated during lagging-strand DNA replication. Efficient nick processing during Okazaki fragment maturation requires the coordinated action of DNA polymerase δ (Pol δ) 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 δ 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 δ 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 ϵ is the leading-strand enzyme, we observed no idling by this enzyme and no cooperation with FEN1 for creating a ligatable nick.
Keyword
- DNA/*metabolism
- DNA Helicases/metabolism
- DNA Polymerase III/*metabolism
- DNA Primers
- DNA Replication
- Exonucleases/metabolism
- Flap Endonucleases/*metabolism
- Oligonucleotides/metabolism
- Saccharomyces cerevisiae/*enzymology/*genetics
Publication and Content Type
- ref (subject category)
- art (subject category)
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