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J Bacteriol. 1971 October; 108(1): 364-374
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Altered Deoxyribonucleic Acid Polymerase Activity in a Methyl Methanesulfonate-Sensitive Mutant of Bacillus subtilis

Kenneth B. Gass, Thomas C. Hill, Mehran Goulian1, Bernard S. Strauss and Nicholas R. Cozzarelli

a Departments of Biochemistry and Microbiology, University of Chicago, Chicago, Illinois 60637

ABSTRACT

Of six deoxyribonucleic acid repair mutants of Bacillus subtilis assayed for deoxyribonucleic acid polymerase, only the methyl methanesulfonate-sensitive and ultraviolet light-sensitive mutant JB1-49(59) has impaired polymerase activity. Extracts prepared by sonic treatment or gentle lysis had about 10% of the wild-type activity with poly d(A–T), an alternating copolymer of deoxyadenylate and deoxythymidylate, used as template. The sensitivity to methyl methanesulfonate and ultraviolet light and the low level of polymerase activity transformed and reverted together, indicating that the two characteristics are a pleiotropic manifestation of a single mutation. Mixed extract and kinetic experiments mitigated against an altered nuclease activity as the enzymatic consequence of the mutation. Also, the mutant and wild type activities were stimulated equally by Escherichia coli exonuclease III. The residual activity in the mutant showed several differences from the wild-type activity: it purified differently, was more sensitive to sulfhydryl reagents, and displayed a different template specificity. We tentatively conclude that either the mutation in JB1-49(59) has introduced a qualitative as well as a quantitative change in the polymerase or the wild type contains two distinct polymerases, one of which is missing in the mutant.


FOOTNOTES

1 Present address: Department of Medicine, University of California at San Diego, La Jolla, Calif. 92037.


J Bacteriol. 1971 October; 108(1): 364-374
Copyright © 1971 American Society for Microbiology. All Rights Reserved.




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