Journal of Bacteriology, January 1999, p. 204-211, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
E Is Required for Normal Cell Wall Structure
in Streptomyces coelicolor A3(2)
John Innes Centre,
Received 28 August 1998/Accepted 20 October 1998
The sigE gene of Streptomyces coelicolor
A3(2) encodes an RNA polymerase sigma factor belonging to the
extracytoplasmic function (ECF) subfamily. Constructed sigE
deletion and disruption mutants were more sensitive than the parent to
muramidases such as hen egg white lysozyme and to the CwlA amidase from
Bacillus subtilis. This correlated with an altered
muropeptide profile, as determined by reverse-phase high-performance
liquid chromatography analysis of lytic digests of purified
peptidoglycan. The sigE mutants required high levels of
magnesium for normal growth and sporulation, overproducing the
antibiotic actinorhodin and forming crenellated colonies in its
absence. Together, these data suggest that sigE is required for normal cell wall structure. The role of
E was
further investigated by analyzing the expression of hrdD, which is partially sigE dependent. The hrdD
gene, which encodes the
HrdD subunit of RNA polymerase,
is transcribed from two promoters, hrdDp1 and
hrdDp2, both similar to promoters recognized by
other ECF sigma factors. The activities of
hrdDp1 and hrdDp2 were
reduced 20- and 3-fold, respectively, in sigE mutants,
although only hrdDp1 was recognized by
E
E in vitro. Growth on media deficient in magnesium
caused the induction of both hrdD promoters in a
sigE-dependent manner.
*
Corresponding author. Mailing address: John Innes
Centre, Colney, Norwich NR4 7UH, United Kingdom. Phone: (44) 1603 452571. Fax: (44) 1603 456844. E-mail:
pagetm{at}bbsrc.ac.uk.
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