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Volume 270,
Number 31,
Issue of August 04, pp. 18335-18340, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Effects
of the Antiterminator BoxA on Transcription Elongation Kinetics and
ppGpp Inhibition of Transcription Elongation in Escherichia coli
(Received for publication, March 9, 1995; and in revised form, May 12, 1995)
Ulla
Vogel
,
Kaj
Frank
Jensen
It has been shown previously that two different mRNA chains (lacZ and infB) are elongated at a rate of 40
nucleotides (nt)/s during steady state growth on minimal medium and
that the rate of mRNA chain elongation is inhibited by ppGpp in
vivo. On the other hand, it was found that a truncated ribosomal
RNA chain was elongated at a rate of 80 nt/s, independent of
growth condition (Vogel, U., and Jensen, K. F. (1994) J. Biol.
Chem. 269, 16236-16241). We reasoned that the different
transcriptional behavior of mRNA genes and rRNA operons might be caused
by the antiterminator sequences present in the rRNA operons. To test
this possibility, we have (a) inserted the minimal
antiterminator boxA sequence between the promoter and the lacZ and infB genes and (b) deleted the
antiterminator sequences from the rRNA transcription unit and measured
transcription elongation rates in vivo on the resulting hybrid
genes. We found that insertion of boxA in front of the coding region of lacZ increased the transcription elongation rate from 42 nt/s
to 69 nt/s during steady state growth and that it eliminated the
ppGpp-dependent decrease in the transcription elongation rate during
the stringent response. On the other hand, deletion of the
antiterminator sequences from the rRNA operon resulted in a reduced
transcription elongation rate, but the elongation rate was still
insensitive to changes in the ppGpp pool. These results are consistent
with the hypothesis that the antiterminator boxA is a primary
determinant of the rate of transcription elongation rate.

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Copyright © 1995 by the American Society for Biochemistry and Molecular Biology.
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