A UNIQUE SUBPOPULATION OF MURINE DNA-POLYMERASE ALPHA/PRIMASE SPECIFICALLY INTERACTS WITH POLYOMAVIRUS T-ANTIGEN AND STIMULATES DNA-REPLICATION

被引:18
作者
MOSES, K [1 ]
PRIVES, C [1 ]
机构
[1] COLUMBIA UNIV,DEPT BIOL SCI,NEW YORK,NY 10027
关键词
D O I
10.1128/MCB.14.4.2767
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Murine cells or cell extracts support the replication of plasmids containing the replication origin (ori-DNA) of polyomavirus (Py) but not that of simian virus 40 (SV40), whereas human cells or cell extracts support the replication of SV40 ori-DNA but not that of Py ori-DNA. It was shown previously that fractions containing DNA polymerase alpha/primase from permissive cells allow viral ori-DNA replication to proceed in extracts of nonpermissive cells. To extend these observations, the binding of Py T antigen to both the permissive and nonpermissive DNA polymerase alpha/primase was examined. Py T antigen was retained by a murine DNA polymerase alpha/primase but not by a human DNA polymerase alpha/primase affinity column. Likewise, a Py T antigen affinity column retained DNA polymerase alpha/primase activity from murine cells but not from human cells. The murine fraction which bound to the Py T antigen column was able to stimulate Py ori-DNA replication in the nonpermissive extract. However, the DNA polymerase alpha/primase activity in this murine fraction constituted only a relatively small proportion (approximately 20 to 40%) of the total murine DNA polymerase alpha/primase that had been applied to the column. The DNA polymerase alpha/primase purified from the nonbound murine fraction, although far more replete in this activity, was incapable of supporting Py DNA replication. The two forms of murine DNA polymerase alpha/primase also differed in their interactions with Py T antigen. Our data thus demonstrate that there are two distinct populations of DNA polymerase alpha/primase in murine cells and that species-specific interactions between T antigen and DNA polymerases can be identified. They may also provide the basis for initiating a novel means of characterizing unique subpopulations of DNA polymerase alpha/primase.
引用
收藏
页码:2767 / 2776
页数:10
相关论文
共 59 条
[41]   EXPRESSION OF POLYOMAVIRUS LARGE T-ANTIGEN BY USING A BACULOVIRUS VECTOR [J].
RICE, WC ;
LORIMER, HE ;
PRIVES, C ;
MILLER, LK .
JOURNAL OF VIROLOGY, 1987, 61 (05) :1712-1716
[42]   SIMIAN VIRUS-40 AND POLYOMAVIRUS LARGE TUMOR-ANTIGENS HAVE DIFFERENT REQUIREMENTS FOR HIGH-AFFINITY SEQUENCE-SPECIFIC DNA-BINDING [J].
SCHELLER, A ;
PRIVES, C .
JOURNAL OF VIROLOGY, 1985, 54 (02) :532-545
[43]   DNA HELICASE AND NUCLEOSIDE-5'-TRIPHOSPHATASE ACTIVITIES OF POLYOMA-VIRUS LARGE TUMOR-ANTIGEN [J].
SEKI, M ;
ENOMOTO, T ;
EKI, T ;
MIYAJIMA, A ;
MURAKAMI, Y ;
HANAOKA, F ;
UI, M .
BIOCHEMISTRY, 1990, 29 (04) :1003-1009
[44]   T-ANTIGEN-DNA POLYMERASE-ALPHA COMPLEX IMPLICATED IN SIMIAN VIRUS-40 DNA-REPLICATION [J].
SMALE, ST ;
TJIAN, R .
MOLECULAR AND CELLULAR BIOLOGY, 1986, 6 (11) :4077-4087
[45]   DNA HELICASE ACTIVITY OF SV40 LARGE TUMOR-ANTIGEN [J].
STAHL, H ;
DROGE, P ;
KNIPPERS, R .
EMBO JOURNAL, 1986, 5 (08) :1939-1944
[46]  
STILLMAN B, 1989, ANNU REV CELL BIOL, V5, P197
[47]  
SUZUKI M, 1989, J BIOL CHEM, V264, P10065
[48]  
TANAKA S, 1982, J BIOL CHEM, V257, P8386
[49]  
Tooze J, 1981, MOL BIOL TUMOR VIRUS
[50]   SEQUENTIAL INITIATION OF LAGGING AND LEADING STRAND SYNTHESIS BY 2 DIFFERENT POLYMERASE COMPLEXES AT THE SV40 DNA-REPLICATION ORIGIN [J].
TSURIMOTO, T ;
MELENDY, T ;
STILLMAN, B .
NATURE, 1990, 346 (6284) :534-539