Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Insight into the Mechanism of the Stabilization of Moloney Murine Leukaemia Virus Reverse Transcriptase by Eliminating RNase H Activity
Masaki MIZUNOKiyoshi YASUKAWAKuniyo INOUYE
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JOURNALS FREE ACCESS Advance online publication

Article ID: 90777

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Abstract

We explored the mechanism of the stabilization of Moloney murine leukaemia virus reverse transcriptase (MMLV RT) by eliminating RNase H activity. Without the template-primer (T/P) poly(rA)-p(dT)15, the temperature reducing initial reverse-transcription activity by 50% over a 10-min incubation of the RNase H activity-deficient variant D524A was higher by 3.7 °C than that of the wild-type enzyme (WT). In the reverse transcription reaction, the Km values for T/P of WT and D524A were almost the same. These results suggest that elimination of RNase H activity enhanced the intrinsic thermal stability of MMLV RT rather than its affinity toward T/P.

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© 2010 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
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