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Journal of Bacteriology, September 1999, p. 5234-5241, Vol. 181, No. 17
Department of Microbiology and Immunology,
University of Maryland School of Medicine, Baltimore, Maryland
21201,1 and Division of Bacterial
Products, Center for Biologics Evaluation and Research, Food and
Drug Administration, Bethesda, Maryland 208922
Received 29 March 1999/Accepted 25 June 1999
Bordetella pertussis, the causative agent of whooping
cough, regulates expression of its virulence factors via a
two-component signal transduction system encoded by the bvg
regulatory locus. It has been shown by activation kinetics that several
of the virulence factors are differentially regulated. fha
is transcribed at 10 min following an inducing signal, while
ptx is not transcribed until 2 to 4 h after the
inducing signal. We present data indicating that prn is
transcribed at 1 h, an intermediate time compared to those of
fha and ptx. We have identified
cis-acting sequences necessary for expression of
prn in B. pertussis by using
prn-lac fusions containing alterations in the sequence
upstream of the prn open reading frame. In vitro
transcription and DNase I footprinting analyses provided evidence to
support our hypothesis that BvgA binds to this sequence upstream of
prn to activate transcription from the promoter. Our
genetic data indicate that the region critical for prn
activation extends upstream to position
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Analysis of BvgA Activation of the Pertactin Gene
Promoter in Bordetella pertussis
84. However, these data do
not support the location of the prn transcription start
site as previously published. We used a number of methods, including
prn-lac fusions, reverse transcriptase PCR, and 5' rapid amplification of cDNA ends, to localize and identify the
bvg-dependent 5' end of the prn transcript to
the cytosine at
125 with respect to the published start site.
*
Corresponding author. Mailing address: University Of
Maryland School of Medicine, Department of Microbiology and Immunology, BRB 13-009, 655 W. Baltimore St., Baltimore, MD 21201-1559. Phone: (410) 706-7677. Fax: (410) 706-2129. E-mail:
ncarbone{at}umaryland.edu.
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