title: Fate of direct and inverted repeats in the RNA hypermutagenesis reaction.
abstract: RNA hypermutagenesis results from cDNA synthesis in the presence of highly biased dNTP precursor concentrations and preferentially exploits human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. Such reaction conditions slow down DNA synthesis, which might be conducive to strand transfer and deletion. This has been investigated. A 6 bp inverted repeat nested between 10 bp repeats was efficiently deleted at dCTP concentrations typically used. Inter- or intramolecular strand transfer between 10 bp repeated sequences separated by runs of templated G residues occurred, but at lower concentrations. If RNA hypermutagenesis of a sequence containing direct and inverted repeats is unavoidable, avian myeloblastosis virus (AMV) reverse transcriptase could be used, as strand transfer occurs with much diminished dCTP substrate dependence.
authors.affiliation: Unité de Rétrovirologie Moléculaire, Institute Pasteur, Paris, France.
authors.forename: V
authors.initials: V
authors.lastname: Pezo
authors.forename: M A
authors.initials: MA
authors.lastname: Martinez
authors.forename: S
authors.initials: S
authors.lastname: Wain-Hobson
mesh_terms: D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000096:biosynthesis; D003845:Deoxycytosine Nucleotides; D054303:HIV Reverse Transcriptase; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics*; D009994:Osmolar Concentration; D012313:RNA / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D012190:Retroviridae / Q000201:enzymology; D017384:Sequence Deletion
chemical_list: D018076:DNA, Complementary; D003845:Deoxycytosine Nucleotides; D012313:RNA; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase