Two DNA polymerase sliding clamps from the thermophilic archaeon Sulfolobus solfataricus

被引:48
作者
De Felice, M
Sensen, CW
Charlebois, RL
Rossi, M
Pisani, FM
机构
[1] CNR, Ist Biochim Prot & Enzimol, I-80125 Naples, Italy
[2] NRC, Inst Marine Biosci, Halifax, NS B3H 3Z1, Canada
[3] Univ Ottawa, Canadian Inst Adv Res, Ottawa, ON K1N 6N5, Canada
[4] Univ Naples Federico II, Dipartimento Chim Organ & Biol, I-80134 Naples, Italy
关键词
DNA polymerase; sliding clamp; thermostability; Archaea; DNA replication;
D O I
10.1006/jmbi.1999.2939
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
Herein, we report the identification and characterization of two DNA polymerase processivity factors from the thermoacidophilic archaeon Sulfolobus solfataricus. They, referred to as 039p (244 amino acid residues, 27 kDa) and 048p (249 amino acid residues, 27 kDa), present significant primary structure similarity to eukaryotic proliferating cell nuclear antigen (PCNA). We demonstrate that both 039p and 048p form oligomers in solution and are able to substantially activate the synthetic activity of the single-subunit family B DNA polymerase from S, solfataricus (Sso DNA pol B1) on poly(dA)-oligo(dT) as a primer-template. This stimulatory effect is the result of enhanced DNA polymerase processivity, as indicated by the analysis of the elongation products on polyacrylamide gels. Activation of Sso DNA pol B1 synthetic activity was also observed on linear primed single-stranded M13 mp18 DNA as a template. By immunoblot analysis using specific rabbit antisera, 039p and 048p were both detected in the logarithmic and stationary phases of S. solfataricus growth curve. This is the first report of the identification and biochemical characterization of two distinct DNA polymerase processivity factors from the same organism. The significance of these findings for the understanding of the DNA replication process in Archaea is discussed. (C) 1999 Academic Press.
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页码:47 / 57
页数:11
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