NDLI: Functional Domains of the TOL Plasmid Transcription Factor XylS

Content Provider	PubMed Central
Author	Kaldalu, Niilo Toots, Urve Lorenzo, Victor De Mart, Ustav
Copyright Year	2000
Abstract	The alkylbenzoate degradation genes of Pseudomonas putida TOL plasmid are positively regulated by XylS, an AraC family protein, in a benzoate-dependent manner. In this study, we used deletion mutants and hybrid proteins to identify which parts of XylS are responsible for the DNA binding, transcriptional activation, and benzoate inducibility. We found that a 112-residue C-terminal fragment of XylS binds specifically to the Pm operator in vitro, protects this sequence from DNase I digestion identically to the wild-type (wt) protein, and activates the Pm promoter in vivo. When overexpressed, that C-terminal fragment could activate transcription as efficiently as wt XylS. All the truncations, which incorporated these 112 C-terminal residues, were able to activate transcription at least to some extent when overproduced. Intactness of the 210-residue N-terminal portion was found to be necessary for benzoate responsiveness of XylS. Deletions in the N-terminal and central regions seriously reduced the activity of XylS and caused the loss of effector control, whereas insertions into the putative interdomain region did not change the basic features of the XylS protein. Our results confirm that XylS consists of two parts which probably interact with each other. The C-terminal domain carries DNA-binding and transcriptional activation abilities, while the N-terminal region carries effector-binding and regulatory functions.
Starting Page	1118
File Format	PDF
ISSN	10985530
e-ISSN	10985530
Journal	Journal of Bacteriology
Issue Number	4
Volume Number	182
Language	English
Publisher	American Society for Microbiology
Publisher Date	2000-02-01
Access Restriction	Open
Rights Holder	American Society for Microbiology
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Microbiology

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Molecular cloning of gene xylS of the TOL plasmid: evidence for positive regulation of the xylDEGF operon by xylS.

Activation and Repression of Transcription at the Double Tandem Divergent Promoters for the xylR and xylS Genes of the TOL Plasmid of Pseudomonas putida

Overproduction of the xylS gene product and activation of the xylDLEGF operon on the TOL plasmid.

The TACAN4TGCA motif upstream from the -35 region in the sigma70-sigmaS-dependent Pm promoter of the TOL plasmid is the minimum DNA segment required for transcription stimulation by XylS regulators.

Mutations leading to constitutive expression from the TOL plasmid meta-cleavage pathway operon are located at the C-terminal end of the positive regulator protein XylS.

Upstream binding sequences of the XylR activator protein and integration host factor in the xylS gene promoter region of the Pseudomonas TOL plasmid.

Identification of critical amino-terminal regions of XylS. The positive regulator encoded by the TOL plasmid.

Expression of the regulatory gene xylS on the TOL plasmid is positively controlled by the xylR gene product.

Expression of the TOL plasmid xylS gene in Pseudomonas putida occurs from a alpha 70-dependent promoter or from alpha 70- and alpha 54-dependent tandem promoters according to the compound used for growth.

Functional Domains of the TOL Plasmid Transcription Factor XylS

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Molecular cloning of gene xylS of the TOL plasmid: evidence for positive regulation of the xylDEGF operon by xylS.

Activation and Repression of Transcription at the Double Tandem Divergent Promoters for the xylR and xylS Genes of the TOL Plasmid of Pseudomonas putida

Overproduction of the xylS gene product and activation of the xylDLEGF operon on the TOL plasmid.

The TACAN4TGCA motif upstream from the -35 region in the sigma70-sigmaS-dependent Pm promoter of the TOL plasmid is the minimum DNA segment required for transcription stimulation by XylS regulators.

Mutations leading to constitutive expression from the TOL plasmid meta-cleavage pathway operon are located at the C-terminal end of the positive regulator protein XylS.

Upstream binding sequences of the XylR activator protein and integration host factor in the xylS gene promoter region of the Pseudomonas TOL plasmid.

Identification of critical amino-terminal regions of XylS. The positive regulator encoded by the TOL plasmid.

Expression of the regulatory gene xylS on the TOL plasmid is positively controlled by the xylR gene product.

Expression of the TOL plasmid xylS gene in Pseudomonas putida occurs from a alpha 70-dependent promoter or from alpha 70- and alpha 54-dependent tandem promoters according to the compound used for growth.

Functional Domains of the TOL Plasmid Transcription Factor XylS