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Infection and Immunity, December 1998, p. 5698-5702, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
TetR Is a Positive Regulator of the Tetanus Toxin
Gene in Clostridium tetani and Is Homologous to
BotR
Jean-Christophe
Marvaud,1
Ulrich
Eisel,2
Thomas
Binz,3
Heiner
Niemann,3 and
Michel
R.
Popoff1,*
Unité des Toxines Microbiennes,
Institut Pasteur, 75724 Paris Cedex 15, France,1
and
Institut für Zellbiologie und Immunologie,
University of Stuttgart, 70569 Stuttgart,2
and
Institut für Biochemie, Medizinische Hochschule,
30623 Hannover,3 Germany
Received 6 July 1998/Returned for modification 20 August
1998/Accepted 8 September 1998
The TetR gene immediately upstream from the tetanus toxin (TeTx)
gene was characterized. It encodes a 21,562-Da protein which is related
(50 to 65% identity) to the equivalent genes (botR) in
Clostridium botulinum. TetR has the feature of a DNA
binding protein with a basic pI (9.53). It contains a helix-turn-helix motif and shows 29% identity with other putative regulatory genes in
Clostridium, i.e., uviA from C. perfringens and txeR from C. difficile.
We report for the first time the transformation of C. tetani by electroporation, which permitted us to investigate the
function of tetR. Overexpression of tetR in
C. tetani induced an increase in TeTx production and in the
level of the corresponding mRNA. This indicates that TetR is a
transcriptional activator of the TeTx gene. Overexpression of
botR/A (60% identity with TetR at the amino acid level) in
C. tetani induced an increase in TeTx production comparable
to that for overexpression of tetR. However,
botR/C (50% identity with TetR at the amino acid level) was less efficient. This supports that TetR positively regulates the
TeTx gene in C. tetani and that a conserved mechanism of
regulation of the neurotoxin genes is involved in C. tetani
and C. botulinum.
*
Corresponding author. Mailing address: Unité des
Toxines Microbiennes, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: 33 1 45 68 83 07. Fax: 33 1 45 68 84 56. E-mail: mpopoff{at}pasteur.fr.
Infection and Immunity, December 1998, p. 5698-5702, Vol. 66, No. 12
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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