Structure of Bacterial Regulatory RNAs Determines Their Performance in Competition for the Chaperone Protein Hfq

• Published in: Biochemistry (Easton) Vol. 54; no. 5; pp. 1157 - 1170
• Main Authors: Małecka, Ewelina M, Stróżecka, Joanna, Sobańska, Daria, Olejniczak, Mikołaj
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 10.02.2015
• Subjects: Escherichia coli - chemistry; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - chemistry; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Host Factor 1 Protein - chemistry; Host Factor 1 Protein - genetics; Host Factor 1 Protein - metabolism; Molecular Chaperones - chemistry; Molecular Chaperones - genetics; Molecular Chaperones - metabolism; Mutation; Nucleic Acid Conformation; Protein Binding; RNA, Bacterial - chemistry; RNA, Bacterial - genetics; RNA, Bacterial - metabolism; RNA, Untranslated - chemistry; RNA, Untranslated - genetics; RNA, Untranslated - metabolism
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi500741d

Bacterial regulatory RNAs require the chaperone protein Hfq to enable their pairing to mRNAs. Recent data showed that there is a hierarchy among sRNAs in the competition for access to Hfq, which could be important for the tuning of sRNA-dependent translation regulation. Here, seven structurally different sRNAs were compared using filter-based competition assays. Moreover, chimeric sRNA constructs were designed to identify structure elements important for competition performance. The data showed that besides the 3′-terminal oligouridine sequences also the 5′-terminal structure elements of sRNAs were essential for their competition performance. When the binding of sRNAs to Hfq mutants was compared, the data showed the important role of the proximal and rim sites of Hfq for the binding of six out of seven sRNAs. However, ChiX sRNA, which was the most efficient competitor, bound Hfq in a unique way using the oppositedistal and proximalfaces of this ring-shaped protein. The data indicated that the simultaneous binding to the opposite faces of Hfq was enabled by separate adenosine-rich and uridine-rich sequences in the long, single-stranded region of ChiX. Overall, the results suggest that the individual structural composition of sRNAs serves to tune their performance to different levels resulting in a hierarchy of sRNAs in the competition for access to the Hfq protein.