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Journal of Bacteriology, December 2000, p. 6964-6974, Vol. 182, No. 24
School of Microbiology and Immunology,
University of New South Wales, Sydney, New South Wales 2052, Australia
Received 6 July 2000/Accepted 26 September 2000
We report the cloning, sequencing, and characterization of the
rpoE homolog in Vibrio angustum S14. The
rpoE gene encodes a protein with a predicted molecular mass
of 19.4 kDa and has been demonstrated to be present as a single-copy
gene by Southern blot analysis. The deduced amino acid sequence of RpoE
is most similar to that of the RpoE homolog of Sphingomonas
aromaticivorans,
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Evidence for a Role of rpoE in Stressed
and Unstressed Cells of Marine Vibrio angustum
Strain S14
24, displaying sequence
similarity and identity of 63 and 43%, respectively. Northern blot
analysis demonstrated the induction of rpoE 6, 12, and 40 min after a temperature shift to 40°C. An rpoE mutant was constructed by gene disruption. There was no difference in viability during logarithmic growth, stationary phase, or carbon starvation between the wild type and the rpoE mutant strain. In
contrast, survival of the mutant was impaired following heat shock
during exponential growth, as well as after oxidative stress at 24 h of carbon starvation. The mutant exhibited microcolony formation during optimal growth temperatures (22 to 30°C), and cell area measurements revealed an increase in cell volume of the mutant during
growth at 30°C, compared to the wild-type strain. Moreover, outer
membrane and periplasmic space protein analysis demonstrated many alterations in the protein profiles for the mutant during growth
and carbon starvation, as well as following oxidative stress, in
comparison with the wild-type strain. It is thereby concluded that RpoE
has an extracytoplasmic function and mediates a range of specific
responses in stressed as well as unstressed cells of V. angustum S14.
*
Corresponding author. Mailing address: School of
Microbiology and Immunology, University of New South Wales,
Sydney, NSW 2052, Australia. Phone: 61-2-9385 2102. Fax:
61-2-9385 1779. E-mail: S.Kjelleberg{at}unsw.edu.au.
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