Interaction of tRNA(Lys-3) with multiple forms of human immunodeficiency virus reverse transcriptase.
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, tRNA(Lys-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 reconstitution, 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.
Richter-Cook
N J
NJ
Department of Biochemistry, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106.
Howard
K J
KJ
Cirino
N M
NM
Wöhrl
B M
BM
Le Grice
S F
SF
D002850:Chromatography, Gel; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology* / Q000502:physiology; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D010957:Plasmids; D011485:Protein Binding; D012357:RNA, Transfer, Lys / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D014779:Virus Replication
D046911:Macromolecular Substances; D012357:RNA, Transfer, Lys; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase