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Titolo:
Fidelity of eucaryotic DNA polymerase delta holoenzyme from Schizosaccharomyces pombe
Autore:
Chen, XL; Zuo, SJ; Kelman, Z; ODonnell, M; Hurwitz, J; Goodman, MJ;
Indirizzi:
Univ So Calif, Hedco Mol Biol Labs, Dept Biol Sci & Chem, Los Angeles, CA 90089 USA Univ So Calif Los Angeles CA USA 90089 & Chem, Los Angeles, CA 90089 USA Rockefeller Univ, Howard Hughes Med Inst, New York, NY 10021 USA Rockefeller Univ New York NY USA 10021 s Med Inst, New York, NY 10021 USA Mem Sloan Kettering Canc Ctr, Program Mol Biol, New York, NY 10021 USA MemSloan Kettering Canc Ctr New York NY USA 10021 New York, NY 10021 USA
Titolo Testata:
JOURNAL OF BIOLOGICAL CHEMISTRY
fascicolo: 23, volume: 275, anno: 2000,
pagine: 17677 - 17682
SICI:
0021-9258(20000609)275:23<17677:FOEDPD>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
CELL NUCLEAR ANTIGEN; REPLICATION-FACTOR-C; GEL KINETIC-ANALYSIS; LAGGING-STRAND DNA; ESCHERICHIA-COLI; III HOLOENZYME; SACCHAROMYCES-CEREVISIAE; LEADING-STRAND; SPONTANEOUS MUTATION; ACCESSORY PROTEINS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
48
Recensione:
Indirizzi per estratti:
Indirizzo: Goodman, MJ Univ So Calif, Dept Biol Sci, SHS Rm 172, Los Angeles, CA 90089 USA Univ So Calif SHS Rm 172 Los Angeles CA USA 90089 CA 90089 USA
Citazione:
X.L. Chen et al., "Fidelity of eucaryotic DNA polymerase delta holoenzyme from Schizosaccharomyces pombe", J BIOL CHEM, 275(23), 2000, pp. 17677-17682

Abstract

The fidelity of Schizosaccharomyces pombe DNA polymerase delta was measured in the presence or absence of its processivity subunits, proliferating cell nuclear antigen (PCNA) sliding clamp and replication factor C (RFC) clamp-loading complex, using a synthetic 30-mer primer/100-mer template. Synthesis by pol delta alone was distributive. Processive synthesis occurred in the presence of PCNA, RFC, and Escherichia coli single strand DNA-binding protein (SSB) and required the presence of ATP. "Passive" self-loading of PCNA onto DNA takes place in the absence of RFC, in an ATP-independent reaction, which was strongly inhibited by SSB, The nucleotide substitution error rate for pol delta holoenzyme (HE) (pol delta + PCNA + RFC) was 4.6 x 10(-4)for T.G mispairs, 5.3 x 10(-5) for G.G mispairs, and 4.5 x 10(-6) for A.G mispairs, The T.G misincorporation frequency for pol delta without the accessory proteins was unchanged. The fidelity of pol delta HE was between 1 and 2 orders of magnitude lower than that measured for the E, coli pol III HEat the same template position. This relatively low fidelity was caused by inefficient proofreading by the S, pombe polymerase-associated proofreadingexonuclease, The S. pombe S'-exonuclease activity was also extremely inefficient in excising primer-3'-terminal mismatches in the absence of dNTP substrates and in hydrolyzing single-stranded DNA. A comparison of po; delta HE with E, coli pol III alpha HE (lacking the proofreading exonuclease subunit) showed that both holoenzymes exhibit similar error rates for each mispair.

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Documento generato il 14/07/20 alle ore 05:29:18