title: Gln(84) of moloney murine leukemia virus reverse transcriptase regulates the incorporation rates of ribonucleotides and deoxyribonucleotides.
abstract: Moloney murine leukemia virus reverse transcriptase (RT) selectively uses deoxyribonucleotides over ribonucleotides (rNTPs) as substrates. Substitution of F155 with valine (F155V) was previously found to increase the enzyme's affinity for rNTPs, though without affecting the V(max) for catalysis, and thereby conferred to the enzyme significant RNA polymerase activity. We have sought new mutations that might increase the RNA polymerase activity of the F155V mutant. We report here that substitution of Q84 with alanine improved RT-F155V's RNA polymerase activity, but also its DNA polymerase activity. Kinetic analysis and gel-retardation assays suggested that the substitution increased the enzyme's general affinity for the template-primer.
authors.affiliation: Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Shufeng
authors.initials: S
authors.lastname: Liu
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Goff
authors.forename: Guangxia
authors.initials: G
authors.lastname: Gao
mesh_terms: D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000378:metabolism; D005973:Glutamine; D007700:Kinetics; D008979:Moloney murine leukemia virus / Q000201:enzymology*; D020125:Mutation, Missense; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012265:Ribonucleotides / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D003854:Deoxyribonucleotides; D012265:Ribonucleotides; D005973:Glutamine; D012194:RNA-Directed DNA Polymerase; D012321:DNA-Directed RNA Polymerases