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Titolo:
Analysis of unassisted translesion replication by the DNA polymerase III holoenzyme
Autore:
Tomer, G; Livneh, Z;
Indirizzi:
Weizmann Inst Sci, Fac Biochem, Dept Biol Chem, IL-76100 Rehovot, Israel Weizmann Inst Sci Rehovot Israel IL-76100 Chem, IL-76100 Rehovot, Israel
Titolo Testata:
BIOCHEMISTRY
fascicolo: 18, volume: 38, anno: 1999,
pagine: 5948 - 5958
SICI:
0006-2960(19990504)38:18<5948:AOUTRB>2.0.ZU;2-K
Fonte:
ISI
Lingua:
ENG
Soggetto:
SINGLE-STRANDED-DNA; ULTRAVIOLET-LIGHT MUTAGENESIS; SITE-SPECIFIC MUTAGENESIS; HUMAN CELL-EXTRACTS; ESCHERICHIA-COLI; ABASIC SITE; DEOXYRIBONUCLEIC-ACID; SOS MUTAGENESIS; UV MUTAGENESIS; APURINIC SITES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
57
Recensione:
Indirizzi per estratti:
Indirizzo: Livneh, Z Weizmann Inst Sci, Fac Biochem, Dept Biol Chem, IL-76100 Rehovot, Israel Weizmann Inst Sci Rehovot Israel IL-76100 6100 Rehovot, Israel
Citazione:
G. Tomer e Z. Livneh, "Analysis of unassisted translesion replication by the DNA polymerase III holoenzyme", BIOCHEM, 38(18), 1999, pp. 5948-5958

Abstract

DNA damage-induced mutations are formed when damaged nucleotides present in single-stranded DNA are replicated. We have developed a new method for the preparation of gapped plasmids containing site-specific damaged nucleotides, as model DNA substrates for translesion replication. Using these substrates, we show that the DNA polymerase III holoenzyme from Escherichia coli can bypass a synthetic abasic site analogue with high efficiency (30% bypass in 16 min), unassisted by other proteins. The theta and tau subunits of the polymerase were not essential for bypass. No bypass was observed when the enzyme was assayed on a synthetic 60-mer oligonucleotide carrying the same lesion, and bypass on a linear gapped plasmid was 3-4-fold slower than ona circular gapped plasmid. There was no difference in the bypass when standing-start and running-start replication were compared. A comparison of translesion replication by DNA polymerase I, DNA polymerase II, the DNA polymerase LII core, and the DNA polymerase III holoenzyme clearly showed that the DNA polymerase III holoenzyme was by far the most effective in performingtranslesion replication. This was not only due to the high processivity ofthe pol III holoenzyme, because increasing the processivity of pol II by adding the gamma complex and beta subunit, did not increase bypass. These results support the model that SOS regulation was imposed on a fundamentally constitutive translesion replication reaction to achieve tight control of mutagenesis.

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Documento generato il 05/12/20 alle ore 01:02:21