Expression of enzymatically active reverse transcriptase of simian immunodeficiency virus in bacteria: Sensitivity to nucleotide analogue inhibitors

Vinayaka R. Prasad; Kyl Myrick; William Haseltine; Stephen P. Goff

doi:10.1016/0042-6822(90)90164-M

Expression of enzymatically active reverse transcriptase of simian immunodeficiency virus in bacteria: Sensitivity to nucleotide analogue inhibitors

Vinayaka R. Prasad, Kyl Myrick, William Haseltine, Stephen P. Goff

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Abstract

A fragment of the SIV_mac251 pol gene was expressed in Escherichia coli as a trpE fusion protein. Analysis of extracts from bacteria containing this expression plasmid revealed the presence of a reverse transcriptase activity dependent on Mgt²⁺ as divalent cation and active on both poly(rA) · oligo(dT) and poly(rC · oligo(dG) templates. In comparative studies, the SIV and HIV-1 reverse transcriptases expressed in bacteria displayed very similar high sensitivities to the chain terminator inhibitors AZTTP and ddTTP. The reverse transcriptase of Moloney murine leukemia virus and the DNA polymerase of E. coli were both more resistant to ddTTP, and the E. coli enzyme was significantly more resistant to AZTTP.

Original language	English (US)
Pages (from-to)	896-900
Number of pages	5
Journal	Virology
Volume	179
Issue number	2
DOIs	https://doi.org/10.1016/0042-6822(90)90164-M
State	Published - Dec 1990
Externally published	Yes

ASJC Scopus subject areas

Virology

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/0042-6822(90)90164-M

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title = "Expression of enzymatically active reverse transcriptase of simian immunodeficiency virus in bacteria: Sensitivity to nucleotide analogue inhibitors",

abstract = "A fragment of the SIVmac251 pol gene was expressed in Escherichia coli as a trpE fusion protein. Analysis of extracts from bacteria containing this expression plasmid revealed the presence of a reverse transcriptase activity dependent on Mgt2+ as divalent cation and active on both poly(rA) · oligo(dT) and poly(rC · oligo(dG) templates. In comparative studies, the SIV and HIV-1 reverse transcriptases expressed in bacteria displayed very similar high sensitivities to the chain terminator inhibitors AZTTP and ddTTP. The reverse transcriptase of Moloney murine leukemia virus and the DNA polymerase of E. coli were both more resistant to ddTTP, and the E. coli enzyme was significantly more resistant to AZTTP.",

author = "Prasad, {Vinayaka R.} and Kyl Myrick and William Haseltine and Goff, {Stephen P.}",

note = "Funding Information: Thus work was supported by Publrc Health Service Grant 1 UOl Al 24845 from the National Institute of Allergy and Infectious Diseases to S.P.G. and W.A.H., and by a scholar grant from the American Foundation for AIDS Research to V.R P.",

year = "1990",

month = dec,

doi = "10.1016/0042-6822(90)90164-M",

language = "English (US)",

volume = "179",

pages = "896--900",

journal = "Virology",

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publisher = "Academic Press Inc.",

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TY - JOUR

T1 - Expression of enzymatically active reverse transcriptase of simian immunodeficiency virus in bacteria

T2 - Sensitivity to nucleotide analogue inhibitors

AU - Prasad, Vinayaka R.

AU - Myrick, Kyl

AU - Haseltine, William

AU - Goff, Stephen P.

N1 - Funding Information: Thus work was supported by Publrc Health Service Grant 1 UOl Al 24845 from the National Institute of Allergy and Infectious Diseases to S.P.G. and W.A.H., and by a scholar grant from the American Foundation for AIDS Research to V.R P.

PY - 1990/12

Y1 - 1990/12

N2 - A fragment of the SIVmac251 pol gene was expressed in Escherichia coli as a trpE fusion protein. Analysis of extracts from bacteria containing this expression plasmid revealed the presence of a reverse transcriptase activity dependent on Mgt2+ as divalent cation and active on both poly(rA) · oligo(dT) and poly(rC · oligo(dG) templates. In comparative studies, the SIV and HIV-1 reverse transcriptases expressed in bacteria displayed very similar high sensitivities to the chain terminator inhibitors AZTTP and ddTTP. The reverse transcriptase of Moloney murine leukemia virus and the DNA polymerase of E. coli were both more resistant to ddTTP, and the E. coli enzyme was significantly more resistant to AZTTP.

AB - A fragment of the SIVmac251 pol gene was expressed in Escherichia coli as a trpE fusion protein. Analysis of extracts from bacteria containing this expression plasmid revealed the presence of a reverse transcriptase activity dependent on Mgt2+ as divalent cation and active on both poly(rA) · oligo(dT) and poly(rC · oligo(dG) templates. In comparative studies, the SIV and HIV-1 reverse transcriptases expressed in bacteria displayed very similar high sensitivities to the chain terminator inhibitors AZTTP and ddTTP. The reverse transcriptase of Moloney murine leukemia virus and the DNA polymerase of E. coli were both more resistant to ddTTP, and the E. coli enzyme was significantly more resistant to AZTTP.

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Expression of enzymatically active reverse transcriptase of simian immunodeficiency virus in bacteria: Sensitivity to nucleotide analogue inhibitors

Abstract

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