Fidelity and processivity of Saccharomyces cerevisiae DNA polymerase η

被引:156
作者
Washington, MT [1 ]
Johnson, RE [1 ]
Prakash, S [1 ]
Prakash, L [1 ]
机构
[1] Univ Texas, Med Branch, Sealy Ctr Mol Sci, Galveston, TX 77555 USA
关键词
D O I
10.1074/jbc.274.52.36835
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 yeast RAD30 gene functions in error-free replication of UV-damaged DNA, and RAD30 encodes a DNA polymerase, pol eta, that has the ability to efficiently and correctly replicate past a cis-syn-thymine-thymine dimer in template DNA. To better understand the role of pol eta in damage bypass, we examined its fidelity and processivity on nondamaged DNA templates. Steady-state kinetic analyses of deoxynucleotide incorporation indicate that pol eta has a low fidelity, misincorporating deoxynucleotides with a frequency of about 10(-2) to 10(-3) Also pol eta has a low processivity, incorporating only a Pew nucleotides before dissociating. We suggest that pol eta's low fidelity reflects a flexibility in its active site rendering it more tolerant of DNA damage, while its low processivity limits its activity to reduce errors.
引用
收藏
页码:36835 / 36838
页数:4
相关论文
共 25 条
[1]   Yeast DNA repair proteins Rad6 and Rad18 form a heterodimer that has ubiquitin conjugating, DNA binding, and ATP hydrolytic activities [J].
Bailly, V ;
Lauder, S ;
Prakash, S ;
Prakash, L .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (37) :23360-23365
[2]  
Bambara RA, 1995, METHOD ENZYMOL, V262, P270
[3]   Fidelity of Escherichia coli DNA polymerase III holoenzyme - The effects of beta,gamma complex processivity proteins and epsilon proofreading exonuclease on nucleotide misincorporation efficiencies [J].
Bloom, LB ;
Chen, XL ;
Fygenson, DK ;
Turner, F ;
ODonnell, M ;
Goodman, MF .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (44) :27919-27930
[4]   DETERMINATION OF PYRIMIDINE DIMER UNWINDING ANGLE BY MEASUREMENT OF DNA ELECTROPHORETIC MOBILITY [J].
CIARROCCHI, G ;
PEDRINI, AM .
JOURNAL OF MOLECULAR BIOLOGY, 1982, 155 (02) :177-183
[5]  
Creighton S, 1995, METHOD ENZYMOL, V262, P232
[6]   GEL KINETIC-ANALYSIS OF DNA-POLYMERASE FIDELITY IN THE PRESENCE OF PROOFREADING USING BACTERIOPHAGE-T4 DNA-POLYMERASE [J].
CREIGHTON, S ;
GOODMAN, MF .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (09) :4759-4774
[7]  
ECHOLS H, 1991, ANNU REV BIOCHEM, V60, P477, DOI 10.1146/annurev.biochem.60.1.477
[8]  
Fersht A., 1984, ENZYME STRUCTURE MEC
[9]  
Goodman MF, 1998, GENETICS, V148, P1475
[10]   BIOCHEMICAL BASIS OF DNA-REPLICATION FIDELITY [J].
GOODMAN, MF ;
CREIGHTON, S ;
BLOOM, LB ;
PETRUSKA, J .
CRITICAL REVIEWS IN BIOCHEMISTRY AND MOLECULAR BIOLOGY, 1993, 28 (02) :83-126