Expression of the C-terminus of HIV-1 reverse transcriptase p66 and p51 subunits as a single polypeptide with RNase H activity

Roberto Zúñiga; Sonali Sengupta; Christine Snyder; Oscar Leon; Monica J. Roth

doi:10.1093/protein/gzh071

Expression of the C-terminus of HIV-1 reverse transcriptase p66 and p51 subunits as a single polypeptide with RNase H activity

Roberto Zúñiga, Sonali Sengupta, Christine Snyder, Oscar Leon, Monica J. Roth

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

The C-terminus of the HIV-1 reverse transcriptase heterodimer was reconstructed into a single polypeptide. The construct encodes the p51 thumb (T) and connection (C) subdomains joined through a linker region to the p66 connection (C) and RNase H (R) domain. The TCCR protein was purified from insoluble fractions of Escherichia coli lysates. The TCCR construct maintains Mn²⁺-dependent RNase H activity and specifically cleaves the substrate mimicking the tRNA removal required for second-strand transfer reactions.

Original language	English (US)
Pages (from-to)	581-587
Number of pages	7
Journal	Protein Engineering, Design and Selection
Volume	17
Issue number	7
DOIs	https://doi.org/10.1093/protein/gzh071
State	Published - Jul 2004

All Science Journal Classification (ASJC) codes

Biotechnology
Bioengineering
Biochemistry
Molecular Biology

Keywords

C-terminus/HIV-1 reverse transcriptase
RNase H activity
Single polypeptide
p66 and p51 subunits

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10.1093/protein/gzh071

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abstract = "The C-terminus of the HIV-1 reverse transcriptase heterodimer was reconstructed into a single polypeptide. The construct encodes the p51 thumb (T) and connection (C) subdomains joined through a linker region to the p66 connection (C) and RNase H (R) domain. The TCCR protein was purified from insoluble fractions of Escherichia coli lysates. The TCCR construct maintains Mn2+-dependent RNase H activity and specifically cleaves the substrate mimicking the tRNA removal required for second-strand transfer reactions.",

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AU - Zúñiga, Roberto

AU - Sengupta, Sonali

AU - Snyder, Christine

AU - Leon, Oscar

AU - Roth, Monica J.

PY - 2004/7

Y1 - 2004/7

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AB - The C-terminus of the HIV-1 reverse transcriptase heterodimer was reconstructed into a single polypeptide. The construct encodes the p51 thumb (T) and connection (C) subdomains joined through a linker region to the p66 connection (C) and RNase H (R) domain. The TCCR protein was purified from insoluble fractions of Escherichia coli lysates. The TCCR construct maintains Mn2+-dependent RNase H activity and specifically cleaves the substrate mimicking the tRNA removal required for second-strand transfer reactions.

KW - C-terminus/HIV-1 reverse transcriptase

KW - RNase H activity

KW - Single polypeptide

KW - p66 and p51 subunits

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Expression of the C-terminus of HIV-1 reverse transcriptase p66 and p51 subunits as a single polypeptide with RNase H activity

Abstract

All Science Journal Classification (ASJC) codes

Keywords

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