Point mutations in conserved amino acid residues within the C-terminal domain of HIV-1 reverse transcriptase specifically repress RNase H function

• Published in: FEBS Letters Vol. 257; no. 2; pp. 311 - 314
• Main Authors: Schatz, Octavian, Cromme, Frans V., Grüninger-Leitch, Fiona, Le Grice, Stuart F.J.
• Format: Book Review Journal Article
• Language: English
• Published: England Elsevier B.V 06.11.1989
• Subjects: AIDS/HIV; Amino Acid Sequence; DNA Mutational Analysis; Endoribonucleases - metabolism; HIV-1 - enzymology; HIV-1 - genetics; HIV-1 reverse transcriptase; Molecular Sequence Data; Ribonuclease H; RNA-Directed DNA Polymerase - genetics; RNA-Directed DNA Polymerase - metabolism; RNase H; Site-directed mutagenesis; Structure-Activity Relationship; HIV-1 reverse transcriptase; Site-directed mutagenesis; RNase H
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(89)81559-5

Two single site substitutions (E 478 → Q and H 539 → 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 Ni 2+-nitrilotriacetic acid affinity chromatography. Both enzymes are clearly defective in RNase H function, but exhibit wild type reverse transcriptase activity.