Interaction of tRNA(Lys-3) with multiple forms of human immunodeficiency virus reverse transcriptase

Titelangaben

Richter-Cook, Nancy J. ; Howard, Kathryn J. ; Cirino, Nick M. ; Wöhrl, Birgitta M. ; Le Grice, Stuart F.J.:
Interaction of tRNA(Lys-3) with multiple forms of human immunodeficiency virus reverse transcriptase.
In: The Journal of Biological Chemistry. Bd. 267 (1992) Heft 22 . - S. 15952-15957.
ISSN 1083-351X

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Abstract

The interaction of several forms (p51, p66, and p66/p51) of recombinant human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) with a synthetic derivative of its cognate replication primer, tRNALys-3, has been determined by gel-mobility shift analysis. While p66/p51 RT is proficient in tRNA binding, preparations of p66 and p51 display only weak binding at elevated protein:tRNA ratios, despite the former containing both RNA-dependent DNA polymerase and ribonuclease H (RNase H) activity. Gel permeation analysis of purified p66 RT indicate this to be predominantly monomeric, suggesting that dimerization may be a prerequisite for efficient tRNA binding. Prolonged incubation of a mixture of the 66- and 51- kDa polypeptides results in heterodimer reconstitu- tion, restoration of tRNA binding, and recovery of appreciable levels of RNA-dependent DNA polymerase activity. Under the same conditions, both the tRNA binding and RNA dependent DNA polymerase activities of the 66- and 51-kDa polypeptides are unaffected, suggesting that they remain in the monomeric conformation.