Reverse transcriptases (RTs) from Moloney murine leukaemia virus (MMLV) and avian myeloblastosis virus (AMV) contain all the fingers, palm, thumb, connection and RNase H domains. The fingers, palm and thumb domains are thought to be involved in the reverse transcription activity, and the RNase H domain is in the RNase H activity. In this study, we characterized four chimeric RTs which comprise one of the fingers, palm, thumb and RNase H domains originated from AMV RT and the other three and connection domains originated from MMLV RT. Unexpectedly, all chimeric RTs exhibited the same characteristics: their specific reverse transcription activities decreased to less than 0.1% of that of MMLV RT, while their specific RNase H activities were approximately 20% of that of MMLV RT. The decreases in the two activities of the chimeric RTs were ascribed to the decreases in k(cat). Based on that the reverse transcription activity of MMLV RT was impaired by substituting its RNase H domain with that from AMV RT, we propose that in MMLV RT, there might be an interaction between the fingers/palm/thumb domain and the RNase H domain.
Yasukawa K, Nemoto D, Inouye K. Yasukawa K, et al. J Biochem. 2008 Feb;143(2):261-8. doi: 10.1093/jb/mvm217. Epub 2007 Nov 15. J Biochem. 2008. PMID: 18006517
Gao HQ, Sarafianos SG, Arnold E, Hughes SH. Gao HQ, et al. J Mol Biol. 1999 Dec 17;294(5):1097-113. doi: 10.1006/jmbi.1999.3325. J Mol Biol. 1999. PMID: 10600369
Goedken ER, Marqusee S. Goedken ER, et al. Proteins. 1998 Oct 1;33(1):135-43. Proteins. 1998. PMID: 9741851
Takaku H, Kim SG, Hatta T, Takai K. Takaku H, et al. Nihon Rinsho. 1995 Mar;53(3):771-8. Nihon Rinsho. 1995. PMID: 7535367 Review. Japanese.
Hizi A, Herschhorn A. Hizi A, et al. Virus Res. 2008 Jun;134(1-2):203-20. doi: 10.1016/j.virusres.2007.12.008. Epub 2008 Mar 3. Virus Res. 2008. PMID: 18291546 Review.
Oscorbin IP, Filipenko ML. Oscorbin IP, et al. Comput Struct Biotechnol J. 2021 Nov 22;19:6315-6327. doi: 10.1016/j.csbj.2021.11.030. eCollection 2021. Comput Struct Biotechnol J. 2021. PMID: 34900141 Free PMC article. Review.
Hisayoshi T, Shinomura M, Yokokawa K, Kuze I, Konishi A, Kawaji K, Kodama EN, Hata K, Takahashi S, Nirasawa S, Sakuda S, Yasukawa K. Hisayoshi T, et al. J Nat Med. 2015 Jul;69(3):432-40. doi: 10.1007/s11418-015-0885-9. Epub 2015 Feb 8. J Nat Med. 2015. PMID: 25663480