translation agency

Cleavage of the HIV-1 p66 reverse transcriptase/RNase H by the p9 protease in vitro generates active p15 RNase H.
Schulze T; Nawrath M; Moelling K; Max-Planck-Institut fur Molekulare
October 30, 1991
Arch Virol. 1991;118(3-4):179-88. Unique Identifier : AIDSLINE

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.

Chromatography, High Pressure Liquid Endoribonucleases/*METABOLISM Gene Products, pol/*METABOLISM HIV Protease/METABOLISM HIV-1/*ENZYMOLOGY Oligopeptides/CHEMICAL SYNTHESIS/METABOLISM RNA-Directed DNA Polymerase/*METABOLISM Sodium Dodecyl Sulfate/PHARMACOLOGY Support, Non-U.S. Gov't JOURNAL ARTICLE