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The Crc and Hfq proteins of Pseudomonas putida cooperate in catabolite repression and formation of ribonucleic acid complexes with specific target motifs
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2015 (English)In: Environmental Microbiology, ISSN 1462-2912, E-ISSN 1462-2920, Vol. 17, no 1, 105-118 p.Article in journal (Refereed) Published
Abstract [en]

The Crc protein is a global regulator that has a key role in catabolite repression and optimization of metabolism in Pseudomonads. Crc inhibits gene expression post-transcriptionally, preventing translation of mRNAs bearing an AAnAAnAA motif [the catabolite activity (CA) motif] close to the translation start site. Although Crc was initially believed to bind RNA by itself, this idea was recently challenged by results suggesting that a protein co-purifying with Crc, presumably the Hfq protein, could account for the detected RNA-binding activity. Hfq is an abundant protein that has a central role in post-transcriptional gene regulation. Herein, we show that the Pseudomonas putidaHfq protein can recognize the CA motifs of RNAs through its distal face and that Crc facilitates formation of a more stable complex at these targets. Crc was unable to bind RNA in the absence of Hfq. However, pull-down assays showed that Crc and Hfq can form a co-complex with RNA containing a CA motif in vitro. Inactivation of the hfq or the crc gene impaired catabolite repression to a similar extent. We propose that Crc and Hfq cooperate in catabolite repression, probably through forming a stable co-complex with RNAs containing CA motifs to result in inhibition of translation initiation.

Place, publisher, year, edition, pages
2015. Vol. 17, no 1, 105-118 p.
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
URN: urn:nbn:se:umu:diva-100780DOI: 10.1111/1462-2920.12499ISI: 000349152800010PubMedID: 24803210OAI: diva2:800052
Available from: 2015-04-01 Created: 2015-03-09 Last updated: 2015-08-12Bibliographically approved
In thesis
1. Multiple regulatory inputs for hierarchical control of phenol catabolism by Pseudomonas putida
Open this publication in new window or tab >>Multiple regulatory inputs for hierarchical control of phenol catabolism by Pseudomonas putida
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Metabolically versatile bacteria have evolved diverse strategies to adapt to different environmental niches and respond to fluctuating physico-chemical parameters. In order to survive in soil and water habitats, they employ specific and global regulatory circuits to integrate external and internal signals to counteract stress and optimise their energy status. One strategic endurance mechanism is the ability to choose the most energetically favourable carbon source amongst a number on offer.

Pseudomonas putida strains possess large genomes that underlie much of their ability to use diverse carbon sources as growth substrates. Their metabolic potential is frequently expanded by possession of catabolic plasmids to include the ability to grow at the expense of seemingly obnoxious carbon sources such as phenols. However, this ability comes with a metabolic price tag. Carbon source repression is one of the main regulatory networks employed to subvert use of these expensive pathways in favour of alternative sources that provide a higher metabolic gain. This thesis identifies some of the key regulatory elements and factors used by P. putida to supress expression of plasmid-encoded enzymes for degradation of phenols until they are beneficial.

I first present evidence for a newly identified DNA and RNA motif within the regulatory region of the gene encoding the master regulator of phenol catabolism – DmpR. The former of these motifs functions to decrease the number of transcripts originating from the dmpR promoter, while the latter mediates a regulatory checkpoint for translational repression by Crc – the carbon repression control protein of P. putida. The ability of Crc to form repressive riboprotein complexes with RNA is shown to be dependent on the RNA chaperone protein Hfq – a co-partnership demonstrated to be required for many previously identified Crc-targets implicated in hierarchical assimilation of different carbon sources in P. putida. Finally, I present evidence for a model in which Crc and Hfq co-target multiple RNA motifs to bring about a two-tiered regulation to subvert catabolism of phenols in the face of preferred substrates – one at the level of the regulator DmpR and another at the level of translation of the catabolic enzymes.

Place, publisher, year, edition, pages
Umeå: Umeå University, 2015. 46 p.
P. putida, Phenol catabolism, dmpR, dmp-System, 5’-LR, Carbon Catabolite Repression, Transcriptional & Translational regulation, Crc, CrcZ, CrcY, Hfq
National Category
Cell and Molecular Biology
urn:nbn:se:umu:diva-106878 (URN)978-91-7601-313-7 (ISBN)
Public defence
2015-09-07, E04, Unod R1, Norrlands universitetssjukhus, Umeå, 10:00 (English)
Available from: 2015-08-17 Created: 2015-08-11 Last updated: 2015-08-12Bibliographically approved

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Madhushani, AnjanaShingler, Victoria
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Department of Molecular Biology (Faculty of Science and Technology)
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