1887

Abstract

Members of the CsrA/RsmA family are global regulatory proteins that bind to mRNAs, usually at the ribosome-binding site, to control mRNA translation and stability. Their activity is counteracted by small non-coding RNAs (sRNAs), which offer several binding sites to compete with mRNA binding. The / genes are widespread in prokaryotic chromosomes, although certain phylogenetic groups such as Alphaproteobacteria lack this type of global regulator. Interestingly, a /-like sequence was identified in the replication region of plasmid pMBA19a from the alphaproteobacterium . This -like allele () is 58 % identical to pv. chromosomal and bears an unusual C-terminal extension that may fold into an extra α-helix. Homology-based modelling of RsmA suggests that all key mRNA-binding residues are conserved and correctly positioned in the RNA-binding pocket. In fact, a 1.6 kb fragment from pMBA19a encompassing the locus restored /-dependent phenotypes of / mutants. The functionality of RsmA was confirmed by the gain of control over target ′–′ and ′–′ translational fusions in the same mutant background. The RsmA activity correlated with Western blot detection of the polypeptide. Phenotype and translational fusion data from / mutants expressing RsmX/Y/Z RNAs indicated that RsmA is able to bind these antagonistic sRNAs. In agreement with the latter observation, it was also found that the sRNA RsmY was stabilized by RsmA. Deletion of the C-terminal extra α-helix of RsmA affected its cellular concentration, but increased its relative RNA-binding activity. This is believed to be the first report of the presence and characterization of a functional / homologue in a mobile genetic element.

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2013-02-01
2019-09-15
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