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 -