Insight into the Mechanism of the Stabilization of Moloney Murine Leukaemia Virus Reverse Transcriptase by Eliminating RNase H Activity

@article{Mizuno2010InsightIT,
  title={Insight into the Mechanism of the Stabilization of Moloney Murine Leukaemia Virus Reverse Transcriptase by Eliminating RNase H Activity},
  author={Masaki Mizuno and Kiyoshi Yasukawa and Kuniyo Inouye},
  journal={Bioscience, Biotechnology, and Biochemistry},
  year={2010},
  volume={74},
  pages={440 - 442}
}
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 K m values for T/P of WT and D524A were almost the… 
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26 Citations
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Selective inactivation of M-MuLV RT RNase H activity by site-directed PEGylation: an improved ability to synthesize long cDNA molecules.
Improvement of Moloney murine leukemia virus reverse transcriptase thermostability by introducing a disulfide bridge in the ribonuclease H region.
TLDR
This study attempted to increase MMLV RT thermostability by introducing a disulfide bridge in its RNase H region using site-directed mutagenesis and found A551C/T662C was determined to be the most thermostable variant.
Stabilization of Moloney murine leukemia virus reverse transcriptase by site-directed mutagenesis of surface residue Val433
TLDR
Single variants of Moloney murine leukemia virus reverse transcriptase, V433R and V433K in which a surface hydrophobic residue, Val433, was mutated, retained 55% of initial reverse transcription activity, while the wild-type enzyme retained 17%.
Further increase in thermostability of Moloney murine leukemia virus reverse transcriptase by mutational combination
TLDR
This study attempted to further increase the thermostability of MM3 by introducing positive charges by site-directed mutagenesis at positions that have been implicated in the interaction with template-primer.
Stabilization of human immunodeficiency virus type 1 reverse transcriptase by site-directed mutagenesis
TLDR
The Asp443 → Ala mutation, which abolishes RNase H activity, was introduced into the p66 subunits of HIV-1 M RT and HIV- 1 O RT by site-directed mutagenesis.
Improving the thermal stability of avian myeloblastosis virus reverse transcriptase α-subunit by site-directed mutagenesis
TLDR
A highly stable AMV RT α-subunit is generated by the same mutation strategy as applied to MMLV RT and that positive charges are introduced into RT at positions that have been implicated to interact with T/P by site-directed mutagenesis.
Increase in thermal stability of Moloney murine leukaemia virus reverse transcriptase by site-directed mutagenesis.
Preparation and characterization of the RNase H domain of Moloney murine leukemia virus reverse transcriptase.
Expression of moloney murine leukemia virus reverse transcriptase in a cell-free protein expression system
TLDR
MMLV RT expressed in the cell-free system is indistinguishable from that expressed in E. coli, and in the cDNA synthesis reaction, MM4 exhibited higher thermostability than WT.
Expression of the thermostable Moloney murine leukemia virus reverse transcriptase by silkworm-baculovirus expression system
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References

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TLDR
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TLDR
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TLDR
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TLDR
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TLDR
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TLDR
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TLDR
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