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Titolo:
EFFICIENT REPLICATION OF POLYOMAVIRUS DNA IN A CELL-FREE SYSTEM SUPPLEMENTED WITH ESCHERICHIA-COLI SINGLE-STRANDED-DNA BINDING-PROTEIN, WHICH EXHIBITS SPECIES-SPECIFICITY IN THE REQUIREMENT FOR DNA-POLYMERASE ALPHA-PRIMASE
Autore:
EKI T; HANAOKA F; KOHDA T; SEKI M; UI M; ENOMOTO T;
Indirizzi:
UNIV TOKYO,FAC PHARMACEUT SCI,DEPT PHYSIOL CHEM,BUNKYO KU,7-3-1 HONGOTOKYO 113 JAPAN
Titolo Testata:
Journal of Biochemistry
fascicolo: 2, volume: 118, anno: 1995,
pagine: 435 - 441
SICI:
0021-924X(1995)118:2<435:EROPDI>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
TEMPERATURE-SENSITIVE MUTANT; MOUSE FM3A CELLS; SV40 T-ANTIGEN; LARGE-TUMOR-ANTIGEN; HEAT-LABILE; TSFT20 STRAIN; RESTRICTIVE TEMPERATURE; INVITRO REPLICATION; INITIATION; ORIGIN;
Keywords:
CELL-FREE SYSTEM; DNA POLYMERASE ALPHA-PRIMASE COMPLEX; POLYOMAVIRUS DNA REPLICATION; SINGLE-STRANDED DNA BINDING PROTEIN; TEMPERATURE-SENSITIVE MUTANT;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
47
Recensione:
Indirizzi per estratti:
Citazione:
T. Eki et al., "EFFICIENT REPLICATION OF POLYOMAVIRUS DNA IN A CELL-FREE SYSTEM SUPPLEMENTED WITH ESCHERICHIA-COLI SINGLE-STRANDED-DNA BINDING-PROTEIN, WHICH EXHIBITS SPECIES-SPECIFICITY IN THE REQUIREMENT FOR DNA-POLYMERASE ALPHA-PRIMASE", Journal of Biochemistry, 118(2), 1995, pp. 435-441

Abstract

We established a modified cell-free system for polyomavirus (PyV) DNAreplication, which was supplemented with Escherichia coli single-stranded DNA binding protein (SSB), DNA synthesis in this system was enhanced by 1.4- to over 15-fold depending upon the amount of cell extractscontained in the reaction mixture, By supplementing with E, coli SSB,we were able to reduce the amount of cell extracts in the reaction mixture, and to lower the concentrations of creatine phosphate and Tris,rendering this system more resistant to salts than the conventional PyV DNA replication system, The modified system was characterized usingmutant cell extracts which had heat-inactivated DNA polymerase alpha. DNA synthesis in the system was dependent on PyV T antigen, the PyV origin of DNA replication, mutant cell extracts, and DNA polymerase alpha-primase complex purified from wild-type cells, The DNA polymerase alpha-primase complex was not replaced by DNA polymerase alpha, indicating that this system requires a functional DNA polymerase alpha-primase complex, This system exhibited species-specificity in the requirement for DNA polymerase alpha-primase; only mouse DNA polymerase alpha-primase but not human DNA polymerase alpha-primase functioned in this system.

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Documento generato il 01/10/20 alle ore 15:31:05