Journal of Biochemistry Advance Access originally published online on November 15, 2007
Journal of Biochemistry 2008 143(2):261-268; doi:10.1093/jb/mvm217
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© 2007 The Japanese Biochemical Society.
Comparison of the Thermal Stabilities of Reverse Transcriptases from Avian Myeloblastosis Virus and Moloney Murine Leukaemia Virus
Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
*To whom correspondence should be addressed. Tel: +81-75-753-6267, Fax: +81-75-753-6265, E-mail: yasukawa{at}kais.kyoto-u.ac.jp
Received October 21, 2007; Accepted November 3, 2007
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Abstract |
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Reverse transcriptases (RTs) from avian myeloblastosis virus (AMV) and Moloney murine leukaemia virus (MMLV) have been most extensively used as a tool for conversion of RNA to DNA. In this study, we compared the thermal stabilities of AMV RT and MMLV RT by observing their irreversible thermal inactivation. The temperatures reducing initial activity by 50% in 10-min incubation, T50, of AMV RT were 47°C without the template-primer (T/P), poly(rA)–p(dT)12–18, and 52°C with the T/P (28 µM). T50 of MMLV RT were 44°C without the T/P and 47°C with the T/P (28 µM). Unexpectedly, AMV RT was considerably activated when incubated with the T/P at 45 and 48°C. Such activation was not observed in MMLV RT. These results suggest that AMV RT and MMLV RT are different in the following: (i) The intrinsic thermal stability of AMV RT is higher than that of MMLV RT; (ii) AMV RT is activated by thermal treatment with the T/P at 45–48°C; and (iii) AMV RT is stabilized by the T/P more potently than MMLV RT. Thermodynamic analysis indicates that thermal inactivation of AMV RT and MMLV RT is due to the large entropy change of activation for thermal inactivation.
Key Words: avian myeloblastosis virus, Moloney murine leukaemia virus, reverse transcriptase, template-primer, thermal stability
Abbreviations: AMV, avian myeloblastosis virus; MMLV, Moloney murine leukaemia virus; RT, reverse transcriptase; T/P, template-primer
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