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Infection and Immunity, October 2000, p. 5575-5580, Vol. 68, No. 10
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Construction and Characterization of a
Mycobacterium tuberculosis Mutant Lacking the Alternate
Sigma Factor Gene, sigF
Ping
Chen,1
Rafael E.
Ruiz,1
Qing
Li,2
Richard F.
Silver,2 and
William R.
Bishai1,3,*
Center for Tuberculosis Research, Department
of International Health, Johns Hopkins University School of Public
Health,1 and Department of Medicine,
Johns Hopkins University School of
Medicine,3 Baltimore, Maryland, and
Department of Medicine, Case Western Reserve University
School of Medicine, Cleveland, Ohio2
Received 20 March 2000/Returned for modification 15 May
2000/Accepted 26 June 2000
The alternate RNA polymerase sigma factor gene, sigF,
which is expressed in stationary phase and under stress conditions in vitro, has been deleted in the virulent CDC1551 strain of
Mycobacterium tuberculosis. The growth rate of the
sigF mutant was identical to that of the isogenic
wild-type strain in exponential phase, although in stationary phase the
mutant achieved a higher density than the wild type. The mutant showed
increased susceptibility to rifampin and rifapentine. Additionally, the
sigF mutant displayed diminished uptake of
chenodeoxycholate, and this effect was reversed by complementation with
a wild-type sigF gene. No differences in short-term
intracellular growth between mutant and wild-type organisms within
human monocytes were observed. Similarly, the organisms did not differ
in their susceptibilities to lymphocyte-mediated inhibition of
intracellular growth. However, mice infected with the
sigF mutant showed a median time to death of 246 days
compared with 161 days for wild-type strain-infected animals
(P < 0.001). These data indicate that M. tuberculosis sigF is a nonessential alternate sigma factor both
in axenic culture and for survival in macrophages in vitro. While the
sigF mutant produces a lethal infection of mice, it is
less virulent than its wild-type counterpart by time-to-death analysis.
*
Corresponding author. Mailing address: Center for
Tuberculosis Research, Johns Hopkins University School of Public
Health, 615 North Wolfe St., Baltimore, MD 21205-2179. Phone: (410)
955-3507. Fax: (410) 614-8173. E-mail: wbishai{at}jhsph.edu.
Infection and Immunity, October 2000, p. 5575-5580, Vol. 68, No. 10
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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