Point mutations in conserved amino acid residues within the C-terminal domain of HIV-1 reverse transcriptase specifically repress RNase H function.
Two single site substitutions (E478----Q and H539----F) were introduced into the C-terminal RNase H domain of HIV-1 reverse transcriptase. These mutant proteins were expressed in Escherichia coli and purified by Ni2+-nitrilotriacetic acid affinity chromatography. Both enzymes are clearly defective in RNase H function, but exhibit wild type reverse transcriptase activity.
Schatz
O
O
Central Research Units, F. Hoffmann-LaRoche Ltd., Basel, Switzerland.
Cromme
F V
FV
Grüninger-Leitch
F
F
Le Grice
S F
SF
D000595:Amino Acid Sequence; D004252:DNA Mutational Analysis; D004722:Endoribonucleases / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D016914:Ribonuclease H; D013329:Structure-Activity Relationship
D012194:RNA-Directed DNA Polymerase; D004722:Endoribonucleases; D016914:Ribonuclease H