Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site

Protein–protein interactions are an important element of signal transfer within and between organisms. They are mainly mediated by short oligopeptide motifs and represent a widely used alternative to small, organic molecules for conveying information. The transcription factor TetR, a regulator of te...

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Main Authors:	Klotzsche, Marcus, Goeke, Dagmar, Berens, Christian, Hillen, Wolfgang
Format:	Online
Language:	English
Published:	Oxford University Press 2007
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1919500/

id	pubmed-1919500
recordtype	oai_dc
spelling	pubmed-19195002007-07-24 Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site Klotzsche, Marcus Goeke, Dagmar Berens, Christian Hillen, Wolfgang Molecular Biology Protein–protein interactions are an important element of signal transfer within and between organisms. They are mainly mediated by short oligopeptide motifs and represent a widely used alternative to small, organic molecules for conveying information. The transcription factor TetR, a regulator of tetracycline resistance in Gram-negative bacteria, is naturally induced by tetracycline or its derivatives. The oligopeptide Tip (Transcription inducing peptide) fused to either N- or C-terminus of Thioredoxin A (TrxA) has been isolated as an artificial inducer for TetR in Escherichia coli. This inducing property can be exploited to monitor the in vivo expression of a protein of interest by fusing Tip to its C-terminus. We improve the induction efficiency of Tip by adding an aromatic amino acid before residue 1 of Tip in C-terminal fusions to TrxA. The induction efficiency of that modified TrxA-Tip fusion is further enhanced when the effector-binding pocket of TetR is enlarged by the N82A and F86A mutations. The double mutant is also insensitive to induction by tetracyclines. Thus, Tip is an exclusive inducer of this TetR variant, representing the first example of fully converting a small molecular weight effector-dependent transcription factor into one depending solely on protein–protein recognition. Oxford University Press 2007-06 2007-06-01 /pmc/articles/PMC1919500/ /pubmed/17545198 http://dx.doi.org/10.1093/nar/gkm357 Text en © 2007 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Klotzsche, Marcus Goeke, Dagmar Berens, Christian Hillen, Wolfgang
spellingShingle	Klotzsche, Marcus Goeke, Dagmar Berens, Christian Hillen, Wolfgang Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site
author_facet	Klotzsche, Marcus Goeke, Dagmar Berens, Christian Hillen, Wolfgang
author_sort	Klotzsche, Marcus
title	Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site
title_short	Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site
title_full	Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site
title_fullStr	Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site
title_full_unstemmed	Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site
title_sort	efficient and exclusive induction of tet repressor by the oligopeptide tip results from co-variation of their interaction site
description	Protein–protein interactions are an important element of signal transfer within and between organisms. They are mainly mediated by short oligopeptide motifs and represent a widely used alternative to small, organic molecules for conveying information. The transcription factor TetR, a regulator of tetracycline resistance in Gram-negative bacteria, is naturally induced by tetracycline or its derivatives. The oligopeptide Tip (Transcription inducing peptide) fused to either N- or C-terminus of Thioredoxin A (TrxA) has been isolated as an artificial inducer for TetR in Escherichia coli. This inducing property can be exploited to monitor the in vivo expression of a protein of interest by fusing Tip to its C-terminus. We improve the induction efficiency of Tip by adding an aromatic amino acid before residue 1 of Tip in C-terminal fusions to TrxA. The induction efficiency of that modified TrxA-Tip fusion is further enhanced when the effector-binding pocket of TetR is enlarged by the N82A and F86A mutations. The double mutant is also insensitive to induction by tetracyclines. Thus, Tip is an exclusive inducer of this TetR variant, representing the first example of fully converting a small molecular weight effector-dependent transcription factor into one depending solely on protein–protein recognition.
publisher	Oxford University Press
publishDate	2007
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1919500/
_version_	1611398279118979072

Efficient and exclusive induction of Tet repressor by the oligopeptide Tip results from co-variation of their interaction site

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