Literature DB >> 1699794

Purification and characterization of the RNase H domain of HIV-1 reverse transcriptase expressed in recombinant Escherichia coli.

S P Becerra1, G M Clore, A M Gronenborn, A R Karlström, S J Stahl, S H Wilson, P T Wingfield.   

Abstract

The ribonuclease H (RNase H) domain of human immuno-deficiency virus (HIV-1) reverse transcriptase has been produced with the aim of providing sufficient amounts of protein for biophysical studies. A plasmid vector is described which directs high level expression of the RNase H domain under the control of the lambda PL promoter. The domain corresponds to residues 427-560 of the 66 kDa reverse transcriptase. The protein was expressed in Escherichia coli and was purified using ion-exchange and size exclusion chromatography. The purified protein appears to be in a native-like homogeneous conformational state as determined by 1H-NMR spectroscopy and circular dichroism measurements. HIV-protease treatment of the RNase H domain resulted in cleavage between Phe-440 and Tyr-441.

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Year:  1990        PMID： 1699794     DOI： 10.1016/0014-5793(90)81238-j

Source DB:  PubMed          Journal:  FEBS Lett        ISSN： 0014-5793            Impact factor:   4.124


  12 in total

1.  Cassette mutagenesis of the reverse transcriptase of human immunodeficiency virus type 1.

Authors:  P L Boyer; A L Ferris; S H Hughes
Journal:  J Virol       Date:  1992-02       Impact factor: 5.103

2.  Characterization of HIV-1 reverse transcriptase with antibodies indicates conformational differences between the RNAse H domains of p 66 and p 15.

Authors:  A M Szilvay; S Nornes; A Kannapiran; B I Haukanes; C Endresen; D E Helland
Journal:  Arch Virol       Date:  1993       Impact factor: 2.574

3.  Fusion with an RNA binding domain to confer target RNA specificity to an RNase: design and engineering of Tat-RNase H that specifically recognizes and cleaves HIV-1 RNA in vitro.

Authors:  Y F Melekhovets; S Joshi
Journal:  Nucleic Acids Res       Date:  1996-05-15       Impact factor: 16.971

4.  Combinatorial selection, inhibition, and antiviral activity of DNA thioaptamers targeting the RNase H domain of HIV-1 reverse transcriptase.

Authors:  Anoma Somasunderam; Monique R Ferguson; Daniel R Rojo; Varatharasa Thiviyanathan; Xin Li; William A O'Brien; David G Gorenstein
Journal:  Biochemistry       Date:  2005-08-02       Impact factor: 3.162

5.  Cleavage of double-stranded RNA by RNase HI from a thermoacidophilic archaeon, Sulfolobus tokodaii 7.

Authors:  Naoto Ohtani; Hiroshi Yanagawa; Masaru Tomita; Mitsuhiro Itaya
Journal:  Nucleic Acids Res       Date:  2004-11-01       Impact factor: 16.971

6.  Purification and characterization of an active human immunodeficiency virus type 1 RNase H domain.

Authors:  J S Smith; M J Roth
Journal:  J Virol       Date:  1993-07       Impact factor: 5.103

7.  Identification of the first archaeal Type 1 RNase H gene from Halobacterium sp. NRC-1: archaeal RNase HI can cleave an RNA-DNA junction.

Authors:  Naoto Ohtani; Hiroshi Yanagawa; Masaru Tomita; Mitsuhiro Itaya
Journal:  Biochem J       Date:  2004-08-01       Impact factor: 3.857

8.  Unfolding the HIV-1 reverse transcriptase RNase H domain--how to lose a molecular tug-of-war.

Authors:  Xunhai Zheng; Lars C Pedersen; Scott A Gabel; Geoffrey A Mueller; Eugene F DeRose; Robert E London
Journal:  Nucleic Acids Res       Date:  2016-01-14       Impact factor: 16.971

9.  Substitution of a highly basic helix/loop sequence into the RNase H domain of human immunodeficiency virus reverse transcriptase restores its Mn(2+)-dependent RNase H activity.

Authors:  J L Keck; S Marqusee
Journal:  Proc Natl Acad Sci U S A       Date:  1995-03-28       Impact factor: 11.205

10.  Contributions of DNA polymerase subdomains to the RNase H activity of human immunodeficiency virus type 1 reverse transcriptase.

Authors:  J S Smith; K Gritsman; M J Roth
Journal:  J Virol       Date:  1994-09       Impact factor: 5.103
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