Affiliation: Kyoto University
Yasukawa K, Mizuno M, Inouye K. Characterization of Moloney murine leukaemia virus/avian myeloblastosis virus chimeric reverse transcriptases. J Biochem. 2009;145:315-24 pubmed publisher
Okano H, Baba M, Hidese R, Iida K, Li T, Kojima K, et al
. Accurate fidelity analysis of the reverse transcriptase by a modified next-generation sequencing. Enzyme Microb Technol. 2018;115:81-85 pubmed publisher
..This is the first report about a precise fidelity monitoring of various RTs by a direct sequence determination. ..
Konishi A, Hisayoshi T, Yokokawa K, Barrioluengo V, MenÃ©ndez Arias L, Yasukawa K. Amino acid substitutions away from the RNase H catalytic site increase the thermal stability of Moloney murine leukemia virus reverse transcriptase through RNase H inactivation. Biochem Biophys Res Commun. 2014;454:269-74 pubmed publisher
..These results suggest that the mutations, E286R, E302K, and L435R increase the thermostability of MMLV RT not by increasing its affinity for T/P but by abolishing its RNase H activity. ..
Nishimura K, Yokokawa K, Hisayoshi T, Fukatsu K, Kuze I, Konishi A, et al
. Preparation and characterization of the RNase H domain of Moloney murine leukemia virus reverse transcriptase. Protein Expr Purif. 2015;113:44-50 pubmed publisher
..The activity levels of WT, Î”C1, and Î”C2 were estimated to be 1%, 0.01%, and 0.01% of full-length MMLV RT activity, indicating that the C helix is important, but not critical, for the activity of the isolated RNase H domain. ..
Katano Y, Hisayoshi T, Kuze I, Okano H, Ito M, Nishigaki K, et al
. Expression of moloney murine leukemia virus reverse transcriptase in a cell-free protein expression system. Biotechnol Lett. 2016;38:1203-11 pubmed publisher
..MMLV RT expressed in the cell-free system is indistinguishable from that expressed in E. coli. ..
Okano H, Katano Y, Baba M, Fujiwara A, Hidese R, Fujiwara S, et al
. Enhanced detection of RNA by MMLV reverse transcriptase coupled with thermostable DNA polymerase and DNA/RNA helicase. Enzyme Microb Technol. 2017;96:111-120 pubmed publisher
..Our results suggest that DNA polymerase with RT activity and DNA/RNA helicase are useful to increase the sensitivity of cDNA synthesis. ..
Yasukawa K, Nemoto D, Inouye K. Comparison of the thermal stabilities of reverse transcriptases from avian myeloblastosis virus and Moloney murine leukaemia virus. J Biochem. 2008;143:261-8 pubmed
..Thermodynamic analysis indicates that thermal inactivation of AMV RT and MMLV RT is due to the large entropy change of activation for thermal inactivation...
Yasukawa K, Iida K, Okano H, Hidese R, Baba M, Yanagihara I, et al
. Next-generation sequencing-based analysis of reverse transcriptase fidelity. Biochem Biophys Res Commun. 2017;492:147-153 pubmed publisher
..Overall, our method could precisely evaluate the fidelity of various RTs with different reaction conditions in a high-throughput manner without the use of expensive optics and troublesome adaptor ligation. ..
Okano H, Baba M, Kawato K, Hidese R, Yanagihara I, Kojima K, et al
. High sensitive RNA detection by one-step RT-PCR using the genetically engineered variant of DNA polymerase with reverse transcriptase activity from hyperthermophilies. J Biosci Bioeng. 2018;125:275-281 pubmed publisher
..Considering that K4polL329A and RTX are stable even at 90-100°C, our results suggest that the one-step RT-PCR with K4polL329A or RTX is more advantageous than the current one. ..