Activity reversal of Tet repressor caused by single amino acid exchanges
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In: Molecular Microbiology, Vol. 53, No. 3, 08.2004, p. 777-789.
Research output: Journal contributions › Journal articles › Research › peer-review
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TY - JOUR
T1 - Activity reversal of Tet repressor caused by single amino acid exchanges
AU - Scholz, Oliver
AU - Henßler, Eva Maria
AU - Bail, Johannes
AU - Schubert, Peter
AU - Bogdanska-Urbaniak, Joanna
AU - Sopp, Sabine
AU - Reich, Marco
AU - Wisshak, Stefanie
AU - Köstner, Martin
AU - Bertram, Ralph
AU - Hillen, Wolfgang
PY - 2004/8
Y1 - 2004/8
N2 - We explore by extensive mutagenesis regions in the sequence allowing reversal of the allosteric response of Tet repressor. The wild type requires anhydrotetracycline for induction. About 100 mutants are presented, which, in contrast, require the drug for repression. Their mutations are clustered at the interface of the DNA- and inducer-binding domains. This interface consists of a central hydrophobic region surrounded by several hydrogen bonds. While most of the mutants described here contain two to five mutations, we found five positions in this region of TetR, at which single amino acid exchanges lead to activity reversal. They may disrupt the hydrogen-bonding network bordering the domain interface. We assume that the mutations cause a repositioning of the DNA reading head with respect to the effector binding core so that the same conformational change can result in opposite activities.
AB - We explore by extensive mutagenesis regions in the sequence allowing reversal of the allosteric response of Tet repressor. The wild type requires anhydrotetracycline for induction. About 100 mutants are presented, which, in contrast, require the drug for repression. Their mutations are clustered at the interface of the DNA- and inducer-binding domains. This interface consists of a central hydrophobic region surrounded by several hydrogen bonds. While most of the mutants described here contain two to five mutations, we found five positions in this region of TetR, at which single amino acid exchanges lead to activity reversal. They may disrupt the hydrogen-bonding network bordering the domain interface. We assume that the mutations cause a repositioning of the DNA reading head with respect to the effector binding core so that the same conformational change can result in opposite activities.
KW - Chemistry
UR - http://www.scopus.com/inward/record.url?scp=3843071122&partnerID=8YFLogxK
U2 - 10.1111/j.1365-2958.2004.04159.x
DO - 10.1111/j.1365-2958.2004.04159.x
M3 - Journal articles
C2 - 15255892
AN - SCOPUS:3843071122
VL - 53
SP - 777
EP - 789
JO - Molecular Microbiology
JF - Molecular Microbiology
SN - 0950-382X
IS - 3
ER -