title: Human immunodeficiency virus reverse transcriptase expressed in transformed yeast cells. Biochemical properties and interactions with bovine tRNALys.
abstract: Human immunodeficiency virus (HIV) reverse transcriptase has been purified from yeast transformed by an autoreplicating plasmid containing the retroviral DNA polymerase gene. The previously described purification procedure for the yeast-expressed reverse transcriptase [Barr, P.J., Power, M.D., Chun Ting Lee-Ng, Gibson, H. & Luciw, P. (1987) Bio/Technology 5, 486-489] has been substantially modified, leading to an increased yield and a higher degree of purity. Several biochemical properties of the enzyme are described (template specificity, effect of DNA synthesis inhibitors); interestingly, HIV reverse transcriptase is highly resistant to N-ethylmaleimide. A complex between the human retroviral enzyme and the bovine tRNALys was shown, using a direct approach, by glycerol gradient centrifugation, as well as by the protective and specific effect of the tRNALys against enzyme inactivation by thermal denaturation and trypsin digestion. A competitive type of inhibition of HIV reverse transcriptase by tRNALys, but not by tRNAVal, is observed when viral RNA or activated DNA are used as templates.
authors: Institut de Biochimie Cellulaire et de Neurochimie du Centre National de la Recherche Scientifique, Bordeaux, France.
authors: M L
authors: ML
authors: Sallafranque-Andreola
authors: D
authors: D
authors: Robert
authors: P J
authors: PJ
authors: Barr
authors: M
authors: M
authors: Fournier
authors: S
authors: S
authors: Litvak
authors: L
authors: L
authors: Sarih-Cottin
authors: L
authors: L
authors: Tarrago-Litvak
mesh_terms: D000818:Animals; D002417:Cattle; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005784:Gene Amplification; D005814:Genes, Viral; D005822:Genetic Vectors; D006678:HIV / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008970:Molecular Weight; D010957:Plasmids; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012357:RNA, Transfer, Lys / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D013698:Templates, Genetic; D014170:Transformation, Genetic; D015678:Viral Structural Proteins / Q000235:genetics*
chemical_list: D012345:RNA, Transfer, Amino Acid-Specific; D012357:RNA, Transfer, Lys; D015678:Viral Structural Proteins; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase