NDLI: Accurate DNA synthesis by Sulfolobus solfataricus DNA polymerase B1 at high temperature

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Acknowledgment to reviewers 2009

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Accurate DNA synthesis by Sulfolobus solfataricus DNA polymerase B1 at high temperature

Content Provider	Springer Nature Link
Author	Zhang, Likui Lou, Huiqiang Guo, Li Zhan, Zhengyan Duan, Zhenhong Guo, Xin Huang, Li
Copyright Year	2009
Abstract	The accuracy of DNA synthesis by DNA polymerase B1 from the hyperthermophilic archaeon Sulfolobus solfataricus (Sso pol B1) at near the physiological temperature was investigated using M13-based mutational assays. Sso pol B1 showed replication fidelity similar to or higher than most viral, bacterial, and eukaryotic replicases. The fidelity of the enzyme was about three times as high at 70°C as at 55°C. Approximately two-thirds of the errors made by the enzyme were single-base substitutions, of which 58% were C → T transition. Frameshift mutations, mostly resulting from single-base deletions, accounted for 19% of the total errors. An exonuclease-deficient mutant of Sso pol B1 was three times as mutagenic as the wild-type enzyme, suggesting that the intrinsic proofreading function contributed only modestly to the fidelity of the enzyme. Kinetic assays showed that the frequencies of all possible misincorporations by an exonuclease-deficient triple-point mutant of Sso pol B1 ranged from 5.4 × 10−5 to 4.6 × 10−4. The high fidelity of this enzyme in DNA synthesis was based primarily on K m difference rather than V max difference. These properties of Sso pol B1 are consistent with the proposed role of the enzyme as a replicase in S. solfataricus.
Starting Page	107
Ending Page	117
Page Count	11
File Format	PDF
ISSN	14310651
Journal	Extremophiles
Volume Number	14
Issue Number	1
e-ISSN	14334909
Language	English
Publisher	Springer Japan
Publisher Date	2009-12-11
Publisher Place	Japan
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Archaea Sulfolobus solfataricus DNA polymerase Fidelity Mutation spectrum Steady-state kinetics Ecology Biotechnology Microbiology
Content Type	Text
Resource Type	Article
Subject	Medicine Molecular Medicine Microbiology

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