Proteins of the RsmA/CsrA family are global translational regulators in many bacterial species. We have determined the solution structure of a complex formed between the RsmE protein, a member of this family from Pseudomonas fluorescens, and a target RNA encompassing the ribosome-binding site of the hcnA gene. The RsmE homodimer with its two RNA-binding sites makes optimal contact with an 5′-A/UCANGGANGU/A-3′ sequence in the mRNA. When tightly gripped by RsmE, the ANGGAN core folds into a loop, favoring the formation of a 3-base-pair stem by flanking nucleotides. We validated these findings by in vivo and in vitro mutational analyses. The structure of the complex explains well how, by sequestering the Shine-Dalgarno sequence, the RsmA/CsrA proteins repress translation.
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We thank C. Dominguez and R. Stefl for help with the structure calculation; K. Starke (Université de Lausanne) for supplying pME6624, pME6629 and pME6638; D. Witmer (ETH Zürich) for assistance in transcribing RNA and S. Auweter, C. Maris, M. Blatter, A. Clery, C. Reimmann and R. Glockshuber for helpful discussions. This investigation was supported by grants from the Swiss National Science Foundation to D.H. and F.H.-T.A., from the Structural Biology National Center of Competence in Research to F.H.-T.A. and I.J. and from the Roche Research Fund for Biology at the ETH Zurich to F.H.-T.A. F.H.-T.A. is a European Molecular Biology Organization Young Investigator.
The authors declare no competing financial interests.
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Schubert, M., Lapouge, K., Duss, O. et al. Molecular basis of messenger RNA recognition by the specific bacterial repressing clamp RsmA/CsrA. Nat Struct Mol Biol 14, 807–813 (2007). https://doi.org/10.1038/nsmb1285
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