title: Expression of mutant alanine tRNAs increases spontaneous mutagenesis in Escherichia coli.
abstract: The expression of mutA, an allele of the glycine tRNA gene glyV, can confer a novel mutator phenotype that correlates with its ability to promote Asp-->Gly mistranslation. Both activities are mediated by a single base change within the anticodon such that the mutant tRNA can decode aspartate codons (GAC/U) instead of the normal glycine codons (GCC/U). Here, we investigate whether specific Asp-->Gly mistranslation is required for the unexpected mutator phenotype. To address this question, we created and expressed 18 individual alleles of alaV, the gene encoding an alanine tRNA, in which the alanine anticodon was replaced with those specifying other amino acids such that the mutant (alaVX) tRNAs are expected to potentiate X-->Ala mistranslation, where X is one of the other amino acids. Almost all alaVX alleles proved to be mutators in an assay that measured the frequency of rifampicin-resistant mutants, with one allele (alaVGlu) being a stronger mutator than mutA. The alaVGlu mutator phenotype resembles that of mutA in mutational specificity (predominantly transversions), as well as SOS independence, but in a puzzling twist differs from mutA in that it does not require a functional recA gene. Our results suggest that general mistranslation (as opposed to Asp-->Gly alone) can induce a mutator phenotype. Furthermore, these findings predict that a large number of conditions that increase translational errors, such as genetic defects in the translational apparatus, as well as environmental and physiological stimuli (such as amino acid starvation or exposure to antibiotics) are likely to activate a mutator response. Thus, both genetic and epigenetic mechanisms can accelerate the acquisition of mutations.
authors.affiliation: Department of Microbiology and Molecular Genetics, UMDNJ - New Jersey Medical School, 185 South Orange Avenue, Newark, NJ 07103, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Dorazi
authors.forename: Josephine J
authors.initials: JJ
authors.lastname: Lingutla
authors.forename: M Zafri
authors.initials: MZ
authors.lastname: Humayun
mesh_terms: D000483:Alleles; D001224:Aspartic Acid / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D004277:DNA, Single-Stranded / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005838:Genotype; D005998:Glycine / Q000235:genetics; D016296:Mutagenesis; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012344:RNA, Transfer, Ala / Q000235:genetics*
chemical_list: D003062:Codon; D017931:DNA Primers; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D012344:RNA, Transfer, Ala; D001224:Aspartic Acid; D005998:Glycine