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RNA Biol. 2013 Jun;10(6):1031-41. doi: 10.4161/rna.24771. Epub 2013 Apr 23.

RNA pentaloop structures as effective targets of regulators belonging to the RsmA/CsrA protein family.

Author information

1
Département de Microbiologie Fondamentale, Université de Lausanne, Bâtiment Biophore, Lausanne, Switzerland. karine.lapouge@unil.ch

Abstract

In the Gac/Rsm signal transduction pathway of Pseudomonas fluorescens CHA0, the dimeric RNA-binding proteins RsmA and RsmE, which belong to the vast bacterial RsmA/CsrA family, effectively repress translation of target mRNAs containing a typical recognition sequence near the translation start site. Three small RNAs (RsmX, RsmY, RsmZ) with clustered recognition sequences can sequester RsmA and RsmE and thereby relieve translational repression. According to a previously established structural model, the RsmE protein makes optimal contacts with an RNA sequence 5'- (A)/(U)CANGGANG(U)/(A)-3', in which the central ribonucleotides form a hexaloop. Here, we questioned the relevance of the hexaloop structure in target RNAs. We found that two predicted pentaloop structures, AGGGA (in pltA mRNA encoding a pyoluteorin biosynthetic enzyme) and AAGGA (in mutated pltA mRNA), allowed effective interaction with the RsmE protein in vivo. By contrast, ACGGA and AUGGA were poor targets. Isothermal titration calorimetry measurements confirmed the strong binding of RsmE to the AGGGA pentaloop structure in an RNA oligomer. Modeling studies highlighted the crucial role of the second ribonucleotide in the loop structure. In conclusion, a refined structural model of RsmE-RNA interaction accommodates certain pentaloop RNAs among the preferred hexaloop RNAs.

KEYWORDS:

CsrA/RsmA; Gac/Rsm pathway; RNA-binding protein; Shine-Dalgarno structure; translational regulation

PMID:
23635605
PMCID:
PMC4111731
DOI:
10.4161/rna.24771
[Indexed for MEDLINE]
Free PMC Article

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