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Nucleic Acids Research, 1988, Vol. 16, No. 16 8129-8146
© 1988

Articles

Mutations within the decoding site of Escherichia coli 16S rRNA: growth rate impairment, lethality and intragenic suppression

Cheryl L. Thomas, Richard J. Gregory*, Gary Winslow+, Akira Muto§ and Robert A. Zimmermann

Department of Biochemistry and Program in Molecular and Cellular Biology, University of Massachusetts Amherst, MA 01003, USA

*Present addresses: Worcester Foundation for Experimental Biology, Shrewsbury, MA 01545

+Present addresses: Departrnent of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, CO 80309, USA

§Present addresses: Laboratory of Molecular Genetics, Department of Biology, Nagoya University, Nagoya 464, Japan

Received June 17, 1988. Accepted July 22, 1988.

Several C->U transitions and small deletions were introduced into the conserved region centered on base C1400 in Escherichia coil 16S rRNA by in vitro mutagenesis. The mutations were placed within rrnB operons on multicopy plasmids under the transcriptional regulation of either the normal rrnB P1 P2 promoters or the temperature-inducible PL promoter from bacteriophage lain and introduced into E. coil hosts. When expressed from the P1P2 promoters, several of the mutant l6S rRNAs impaired cell growth while others, including one in which U replaced C at position 1400 within the ribosoital decoding site, had little or no effect on cell doubling time. However, C->U transitions at positions 1395 and 1407, as well as the deletion of C1400, appeared to render their hosts inviable. Cells in which these mutations were expressed from the AP promoter died within four generations after induction. Unexpectedly, the lethal phenotype was suppressed intragenically by replacement of 01505 with A, C or U. Suppression may alleviate a functional defect in 30S subunits containing the U1395, U1407 or {bigtriangleup}C1400 mutations.


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