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Journal of Bacteriology, October 2001, p. 6095-6106, Vol. 183, No. 20
Department of Molecular Biology, Umeå
University, SE-901 87 Umeå, Sweden
Received 12 March 2001/Accepted 20 July 2001
The RimM protein in Escherichia coli is associated
with free 30S ribosomal subunits but not with 70S ribosomes. A
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.20.6095-6106.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Characterization of Mutations in the
metY-nusA-infB Operon That
Suppress the Slow Growth of a
rimM Mutant
rimM mutant shows a sevenfold-reduced growth rate and
a reduced translational efficiency, probably as a result of aberrant
assembly of the ribosomal 30S subunits. The slow growth and
translational deficiency can be partially suppressed by increased
synthesis of the ribosome binding factor RbfA. Here, we have identified
14 chromosomal suppressor mutations that increase the growth rate of a
rimM mutant by increasing the expression of
rbfA. Nine of these mutations were in the
nusA gene, which is located upstream from
rbfA in the metY-nusA-infB operon; three mutations deleted the transcriptional terminator between infB and rbfA; one was an
insertion of IS2 in infB, creating a new
promoter for rbfA; and one was a duplication, placing a second copy of rbfA downstream from a promoter for the
yhbM gene. Two of the nusA mutations were
identical, while another mutation (nusA98) was identical
to a previously isolated mutation, nusA11, shown to
decrease termination of transcription. The different nusA mutations were found to increase the expression of
rbfA by increasing the read-through of two internal
transcriptional terminators located just downstream from the
metY gene and that of the internal terminator preceding
rbfA. Induced expression of the
nusA+ gene from a plasmid in a
nusA+ strain decreased the read-through of
the two terminators just downstream from metY,
demonstrating that one target for a previously proposed NusA-mediated
feedback regulation of the metY-nusA-infB operon
expression is these terminators. All of the nusA
mutations produced temperature-sensitive phenotypes of
rimM+ strains. The nusA gene
has previously been shown to be essential at 42°C and below 32°C.
Here, we show that nusA is also essential at 37°C.
*
Corresponding author. Mailing address: Department of
Molecular Biology, Umeå University, SE-901 87 Umeå, Sweden.
Phone: 46-90-7856754. Fax: 46-90-772630. E-mail:
Mikael.Wikstrom{at}micro.umu.se.
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