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The Crc global regulator binds to an unpaired Arich motif at the Pseudomonas putida alkS mRNA coding sequence and inhibits translation initiation

The Crc global regulator binds to an unpaired Arich motif at the Pseudomonas putida alkS mRNA coding sequence and inhibits translation initiation,10.1

The Crc global regulator binds to an unpaired Arich motif at the Pseudomonas putida alkS mRNA coding sequence and inhibits translation initiation   (Citations: 9)
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Crc is a key global translational regulator in Pseudomonads that orchestrates the hierarchy of induction of several catabolic pathways for amino acids, sugars, hydrocarbons or aromatic compounds. In the presence of amino acids, which are preferred carbon sources, Crc inhibits transla- tion of the Pseudomonas putida alkS and benR mRNAs, which code for transcriptional regulators of genes required to assimilate alkanes (hydro- carbons) and benzoate (an aromatic compound), respectively. Crc binds to the 50-end of these mRNAs, but the sequence and/or structure recognized, and the way in which it inhibits transla- tion, were unknown. We have determined the sec- ondary structure of the alkS mRNA 50-end through its sensitivity to several ribonucleases and chemical reagents. Footprinting and band-shift assays using variant alkS mRNAs have shown that Crc specifi- cally binds to a short unpaired A-rich sequence located adjacent to the alkS AUG start codon. This interaction is stable enough to prevent for- mation of the translational initiation complex. A similar Crc-binding site was localized at benR mRNA, upstream of the Shine-Dalgarno sequence. This allowed predicting binding sites at other Crc- regulated genes, deriving a consensus sequence that will help to validate new Crc targets and to discriminate between direct and indirect effects of this regulator.
Journal: Nucleic Acids Research - NAR , vol. 37, no. 22, pp. 7678-7690, 2009
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    • ...Crc is an RNAbinding protein (Moreno et al. 2007; Moreno and Rojo 2008) with high affinity for unpaired A-rich sequences (consensus AAXAAXAA where X=C, U or A; Moreno et al. 2009; Sonnleitner et al. 2009)...

    Elisabeth Sonnleitneret al. Small RNAs as regulators of primary and secondary metabolism in Pseudo...

    • ...40

      A master regulator of carbon metabolism in Pseudomonas is an RNA-binding protein named Crc, which, under repressive conditions, inhibits the translation of mRNAs by binding to a sequence just downstream of the AUG start codon (Figure 2)...

    Sergio Sánchezet al. Carbon source regulation of antibiotic production

    • ...The post-transcriptional action of Crc relies on the binding of Crc to an unpaired A-rich motif in the 5’-end of a target mRNA causing inhibition of the initiation of translation [17,18]...
    • ...Based on the size of the Crc product and the proposed mechanism of action, it is thought that Crc binding must occur within -70 to +16 bp relative to the origin of translation [18]...
    • ...There remain, however, very few known direct targets of Crc: only benR [33], alkS [18], xylR and xylB [34] mRNAs from P. putida and amiE mRNA [17] (product of the amidase gene amiE), in P. aeruginosa have been demonstrated to bind Crc...
    • ...Crc binding site detection: estimation of the numbers of Crc regulated genes in Pseudomonas Following the recent discovery that the Crc protein binds to an A-rich motif in the -70 to +16 region of target mRNAs [17,18] a search was performed to catalogue the occurrences of this motif in the forward orientation in the upstream regions of all protein-encoding genes in Pseudomonas strains present in the RSAT database [40] at the time of ...
    • ...These include the three Crc target genes (alkS, benR of P. putida and amiE of P. aeruginosa )t hat have been experimentally shown to bind Crc in the 5’ region of the mRNA [17,18,33]...
    • ...In this case, the interspecific Crc regulation blocks the synthesis of a transcriptional regulator which leads to indirect regulation of the biosynthetic pathway, reminiscent of the cases of alkS and benR in P. putida [18]...

    Patrick Browneet al. Computational prediction of the Crc regulon identifies genus-wide and ...

    • ...In this light, the repression seen for transcription read-through from ORF101284 with glucose and succinate might point to a Crc-type regulation of catabolite repression in Pseudomonas [32,33], although for the time being no specific Crc binding motifs were detected in the ICEclc core region...

    Muriel Gaillardet al. Transcriptome analysis of the mobile genome ICEclc in Pseudomonas knac...

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