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Infection and Immunity, September 2001, p. 5777-5785, Vol. 69, No. 9
Department of Biomedical Sciences, University
at Albany, Albany, New York 12222,1 and
Wadsworth Center, New York State Department of Health,
Albany, New York 12202-20022
Received 15 February 2001/Returned for modification 2 April
2001/Accepted 4 June 2001
The environmental signals that affect gene regulation in
Mycobacterium tuberculosis remain largely unknown
despite their importance to tuberculosis pathogenesis. Other work has
shown that several promoters, including acr (also
known as hspX) (
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.9.5777-5785.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Identification and Characterization of Mycobacterial
Proteins Differentially Expressed under Standing and Shaking
Culture Conditions, Including Rv2623 from a Novel Class of
Putative ATP-Binding Proteins
-crystallin homolog), are
upregulated in shallow standing cultures compared with constantly shaking cultures. Each of these promoters is also induced to a similar
extent within macrophages. The present study used two-dimensional gel
electrophoresis and mass spectrometry to further characterize differences in mycobacterial protein expression during growth under
standing and shaking culture conditions. Metabolic labeling of
M. bovis BCG showed that at least 45 proteins were
differentially expressed under standing and shaking culture conditions.
Rv2623, CysA2-CysA3, Gap, and Acr were identified from each of four
spots or gel bands that were specifically increased in bacteria from standing cultures. An additional standing-induced spot contained two
comigrating proteins, GlcB and KatG. The greatest induction was
observed with Rv2623, a 32-kDa protein of unknown function that was
strongly expressed under standing conditions and absent in shaking
cultures. Analysis using PROBE, a multiple sequence alignment and
database mining tool, classified M. tuberculosis Rv2623
as a member of a novel class of ATP-binding proteins that may be
involved in M. tuberculosis's response to environmental signals. These studies demonstrate the power of combined proteomic and
computational approaches and demonstrate that subtle differences in
bacterial culture conditions may have important implications for the
study of gene expression in mycobacteria.
*
Corresponding author. Mailing address: Wadsworth
Center, New York State Department of Health, P.O. Box 22002, Albany, NY
12202-2002. Phone: (518) 486-4253. Fax: (518) 474-3181. E-mail:
Kathleen.McDonough{at}wadsworth.org.
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