In:
Molecular Microbiology, Wiley, Vol. 53, No. 3 ( 2004-08), p. 777-789
Abstract:
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.
Type of Medium:
Online Resource
ISSN:
0950-382X
,
1365-2958
DOI:
10.1111/mmi.2004.53.issue-3
DOI:
10.1111/j.1365-2958.2004.04159.x
Language:
English
Publisher:
Wiley
Publication Date:
2004
detail.hit.zdb_id:
1501537-3
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