Improving the thermal stability of avian myeloblastosis virus reverse transcriptase α-subunit by site-directed mutagenesis

@article{Konishi2012ImprovingTT,
  title={Improving the thermal stability of avian myeloblastosis virus reverse transcriptase $\alpha$-subunit by site-directed mutagenesis},
  author={Atsushi Konishi and Kiyoshi Yasukawa and Kuniyo Inouye},
  journal={Biotechnology Letters},
  year={2012},
  volume={34},
  pages={1209-1215}
}
Avian myeloblastosis virus reverse transcriptase (AMV RT) is a heterodimer consisting of a 63 kDa α-subunit and a 95 kDa β subunit. Moloney murine leukaemia virus reverse transcriptase (MMLV RT) is a 75 kDa monomer. These two RTs are the most extensively used for conversion of RNA to DNA. We previously developed several mutations that increase the thermostability of MMLV RT and generated a highly stable MMLV RT variant E286R/E302K/L435R/D524A by combining three of them (Glu286→Arg, Glu302→Lys… Expand
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.
TLDR
The 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. Expand
Major groove binding track residues of the connection subdomain of human immunodeficiency virus type 1 reverse transcriptase enhance cDNA synthesis at high temperatures.
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The results demonstrate that the triad of Arg³⁵⁸, Gly, Gly and Ala in the major groove binding track of HIV-1 RT is a major target for RT stabilization, and most relevant for improving reverse transcription efficiency at high temperatures. Expand
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Stabilization of Moloney murine leukemia virus reverse transcriptase by site-directed mutagenesis of surface residue Val433
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Expression of moloney murine leukemia virus reverse transcriptase in a cell-free protein expression system
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The attachment of Sso7d-like protein improves processivity and resistance to inhibitors of M-MuLV reverse transcriptase
TLDR
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TLDR
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Increase in thermal stability of Moloney murine leukaemia virus reverse transcriptase by site-directed mutagenesis.
TLDR
Highly stable MMLV RT variants were generated by site-directed mutagenesis at positions that have been implicated in the interaction with T/P, including Glu286→Ala, Glu302→Lys, and Leu435→Arg, which is known to abolish the RNase H activity and increase the stability. Expand
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TLDR
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TLDR
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