A Little Gene with Big Effects: a serT Mutant Is Defective in flgM Gene Translation

Abstract

ABSTRACT A conditional-lethal mutant was isolated as having a flagellar regulatory phenotype at 30°C and being unable to grow at 42°C. Chromosomal mapping localized the mutation to the serT gene, which encodes an essential serine tRNA species ( \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(\mathbf{tRNA}_{_{cmo}5\mathbf{UGA}}^{Ser}\) \end{document} ). DNA sequence analysis revealed the mutation to be a single base change in G:A at position 10 of the serT gene that lies within the D-stem of the essential \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(\mathbf{tRNA}_{_{cmo}5}\mathbf{UGA}^{Ser}\) \end{document} species. \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(\mathbf{tRNA}_{_{cmo}5}\mathbf{UGA}^{Ser}\) \end{document} recognizes UCA, UCG, and UCU codons, but UCU is also recognized by \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(\mathbf{tRNA_{GGA}^{Ser}}\) \end{document} and UCG by \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(\mathbf{tRNA_{CGA}^{Ser}}\) \end{document} . No other tRNAs are known to read the UCA codon. Thus, the UCA codon is specifically recognized by \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(\mathbf{tRNA}_{_{cmo}5}\mathbf{UGA}^{Ser}\) \end{document} . We show that the anti-σ 28 activity of FlgM is defective in the serT mutant strain. The serT allele causes a 10-fold increase in σ 28 -dependent fliC promoter transcription, indicating a defect in FlgM anti-σ 28 activity in the presence of the serT mutation. The flgM gene contains only one UCA codon. Changing the UCA of flgM to ACG reversed the effect of the serT allele. Implications for context effects in regulation of gene expression are discussed.