Cleavage of the HIV-1 P66 Reverse Transcriptase/RNase H by the P9 Protease in Vitro Generates Active P15 RNase H

Overview

Journal Arch Virol

Specialty Microbiology

Date 1991 Jan 1

PMID 1712581

Citations 6

Authors

T Schulze

M Nawrath

K Moelling

Affiliations

Soon will be listed here.

Abstract

The reverse transcriptase/RNase H of HIV-1 is composed of a p66/p51 heterodimer when analyzed from virus particles. A recombinant reverse transcriptase (RT)/RNase H which after purification consisted mainly of p66 was analyzed as substrate of the purified recombinant HIV-1 protease p9 in vitro. The p66 protein if treated with the protease is processed to a stable p66/p51 heterodimer. A p15 protein is a prominent cleavage product which was identified as the carboxyterminal portion of p66 by means of a monoclonal antibody. It exhibits RNase H activity when tested by activated gel analysis. Presence of SDS during the incubation allowed complete degradation of p66 depending on the conditions, which indicates that conformation of a substrate is relevant for cleavage by the HIV-1 protease. A synthetic heptapeptide AET-FYVD derived from the region between RT and RNase H is cleaved efficiently in vitro by the HIV-1 protease at the F'Y junction, and may mimick a natural cleavage site. P66/p51 heterodimers exhibit higher RT and RNase H activities than p66 when renatured from polyacrylamide gels.

Citing Articles

Structural Maturation of HIV-1 Reverse Transcriptase-A Metamorphic Solution to Genomic Instability.

London R Viruses. 2016; 8(10).

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Structural integrity of the ribonuclease H domain in HIV-1 reverse transcriptase.

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The nature of the N-terminal amino acid residue of HIV-1 RNase H is critical for the stability of reverse transcriptase in viral particles.

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The p66 immature precursor of HIV-1 reverse transcriptase.

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

Szilvay A, Nornes S, Kannapiran A, Haukanes B, Endresen C, Helland D Arch Virol. 1993; 131(3-4):393-403.

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