PURIFICATION AND PROPERTIES OF RNA-DEPENDENT DNA-POLYMERASE FROM CYTOPLASMIC A-TYPE PARTICLES OF MURINE MAMMARY-TUMOR VIRUS

被引:6
作者
KOHNO, M
ISHIHAMA, A
机构
[1] Institute for Virus Research, Kyoto University, Kyoto, 606, Sakyo-ku
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1979年 / 97卷 / 01期
关键词
D O I
10.1111/j.1432-1033.1979.tb13110.x
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
The RNA‐dependent DNA polymerase associated with cytoplasmic A‐type particles of murine mammary tumor virus was isolated to near homogeneity by a procedure which includes dissociation of proteins from RNA by centrifugation in a step gradient of cesium chloride, followed by an affinity chromatography on poly(rC)‐agarose column. Two species of DNA polymerase were separated by the chromatography: enzyme I in 0.55 M NaCl and enzyme II in 0.80 M NaCl eluate, respectively. The purified DNA polymerases consist of two major polypeptides, with molecular weights of 94000 and 42000, as the intrinsic subunits. Both enzyme protomers with a sedimentation coefficient of 6.3–6.4 S and a molecular weight of 115000–120000 associate to form active oligomers in low‐ionic‐strength buffer. Both enzymes catalyzed the hydrolysis of RNA in RNA. DNA hybrids as well as the RNA‐dependent synthesis of DNA; these are the intrinsic activities of the reverse transcriptase from B‐type particles of murine mammary tumor virus as well as from avian and mammalian C‐type oncornaviruses. The general catalytic properties are similar to those of the enzyme from B‐type particles. Compared with DNA polymerases I, DPU'A polymerase II exhibited a high affinity for all the template‐primers tested and, in addition, a high preference for (rC)n· (dG)12–18. Copyright © 1979, Wiley Blackwell. All rights reserved
引用
收藏
页码:257 / 266
页数:10
相关论文
共 29 条
[1]  
BANDYOPADHYAY AK, 1977, J BIOL CHEM, V252, P5883
[2]  
BEERS RF, 1952, J BIOL CHEM, V195, P133
[3]  
BERNHARD W, 1958, CANCER RES, V18, P491
[4]  
CRAVEN GR, 1965, J BIOL CHEM, V240, P2468
[5]   ISOLATION AND CHARACTERIZATION OF RNA-DIRECTED DNA-POLYMERASE FROM A B-TYPE RNA TUMOR-VIRUS [J].
DION, AS ;
VAIDYA, AB ;
FOUT, GS ;
MOORE, DH .
JOURNAL OF VIROLOGY, 1974, 14 (01) :40-46
[6]   RNASE-H AND RNA-DIRECTED DNA-POLYMERASE - ASSOCIATED ENZYMATIC-ACTIVITIES OF MURINE MAMMARY-TUMOR VIRUS [J].
DION, AS ;
WILLIAMS, CJ ;
MOORE, DH .
JOURNAL OF VIROLOGY, 1977, 22 (01) :187-193
[7]   PURIFICATION AND CHARACTERIZATION OF DEOXYRIBONUCLEIC ACID POLYMERASE ASSOCIATED WITH ROUS-SARCOMA VIRUS [J].
FARAS, AJ ;
TAYLOR, JM ;
LEVINSON, WE ;
BISHOP, JM ;
MCDONNELL, JP .
BIOCHEMISTRY, 1972, 11 (12) :2334-+
[8]  
FUKUDA R, 1974, J MOL BIOL, V83, P335
[9]   PURIFICATION AND CHARACTERIZATION OF DNA-POLYMERASE AND RNASE-H-ACTIVITIES IN MOLONEY MURINE SARCOMA-LEUKEMIA VIRUS [J].
GERARD, GF ;
GRANDGENETT, DP .
JOURNAL OF VIROLOGY, 1975, 15 (04) :785-797
[10]   SINGLE SUBUNIT FROM AVIAN MYELOBLASTOSIS VIRUS WITH BOTH RNA-DIRECTED DNA POLYMERASE AND RIBONUCLEASE-H ACTIVITY [J].
GRANDGENETT, DP ;
GERARD, GF ;
GREEN, M .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1973, 70 (01) :230-234