Publication: The CbrB Regulon: Promoter dissection reveals novel insights into the CbrAB expression network in Pseudomonas putida.
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Identifiers
Date
2018-12-17
Authors
Barroso, Rocío
García-Mauriño, Sofía M
Tomás-Gallardo, Laura
Andújar, Eloísa
Pérez-Alegre, Mónica
Santero, Eduardo
Canosa, Inés
Advisors
Journal Title
Journal ISSN
Volume Title
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Abstract
CbrAB is a high ranked global regulatory system exclusive of the Pseudomonads that responds to carbon limiting conditions. It has become necessary to define the particular regulon of CbrB and discriminate it from the downstream cascades through other regulatory components. We have performed in vivo binding analysis of CbrB in P. putida and determined that it directly controls the expression of at least 61 genes; 20% involved in regulatory functions, including the previously identified CrcZ and CrcY small regulatory RNAs. The remaining are porines or transporters (20%), metabolic enzymes (16%), activities related to protein translation (5%) and orfs of uncharacterised function (38%). Amongst the later, we have selected the operon PP2810-13 to make an exhaustive analysis of the CbrB binding sequences, together with those of crcZ and crcY. We describe the implication of three independent non-palindromic subsites with a variable spacing in three different targets; CrcZ, CrcY and operon PP2810-13 in the CbrAB activation. CbrB is a quite peculiar σN-dependent activator since it is barely dependent on phosphorylation for transcriptional activation. With the depiction of the precise contacts of CbrB with the DNA, the analysis of the multimerisation status and its dependence on other factors such as RpoN o IHF, we propose a model of transcriptional activation.
Description
MeSH Terms
Bacterial Proteins
Base Sequence
Binding Sites
Chromatin Immunoprecipitation
Data Mining
Gene Expression Regulation, Bacterial
Models, Biological
Mutagenesis, Site-Directed
Promoter Regions, Genetic
Protein Binding
Pseudomonas putida
RNA, Bacterial
Real-Time Polymerase Chain Reaction
Sequence Analysis, DNA
Transcription Factors
Transcriptional Activation
Base Sequence
Binding Sites
Chromatin Immunoprecipitation
Data Mining
Gene Expression Regulation, Bacterial
Models, Biological
Mutagenesis, Site-Directed
Promoter Regions, Genetic
Protein Binding
Pseudomonas putida
RNA, Bacterial
Real-Time Polymerase Chain Reaction
Sequence Analysis, DNA
Transcription Factors
Transcriptional Activation