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J. Biol. Chem., Vol. 277, Issue 8, 6170-6177, February 22, 2002
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From the Department of Biochemistry, Robert Wood Johnson Medical
School, Piscataway, New Jersey 08854
In vitro
transcription of lonD, a heat-shock gene from
Myxococcus xanthus, was stimulated in the presence of
extract from heat-shocked cells. For this stimulation the upstream
promoter region of lonD was found to be essential.
Activation of lonD transcription was also observed when
extract from non-heat-shocked cells was heat treated in
vitro at 42 °C for 10 min. A DNA binding assay and
footprinting analysis revealed that a factor(s) binds to the upstream
region from The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF285783.
Transcriptional Activation of a Heat-shock Gene,
lonD, of Myxococcus xanthus by a Two Component
Histidine-Aspartate Phosphorelay System*
122 to 107 with respect to the transcription initiation
site. This region was required for heat-shock induction of
lonD expression both in vitro and in
vivo. The lonD promoter-binding protein named HsfA
was purified, and its gene was cloned. Analysis of the DNA sequence
reveals that HsfA is a response regulator of the two-component system
and shows high sequence similarity to the NtrC family or the
enhancer-binding proteins. Upstream of hsfA, a gene
encoding a histidine kinase was identified and named hsfB.
HsfB was found to be autophosphorylated and able to phosphorylate HsfA.
HsfA with HsfB activated in vitro transcription of
lonD in a manner dependent on RNA polymerase containing
SigA, the housekeeping sigma factor of M. xanthus.
*
This work was supported by a grant from the Foundation of
University of Medicine and Dentistry of New Jersey.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Biochemistry, Robert Wood Johnson Medical School, 675 Hoes La.,
Piscataway, NJ 08854. Tel.: 732-235-4161; Fax: 732-235-4559;
E-mail; sinouye@waksman.rutgers.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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