Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663)

• Published in: RNA (Cambridge) 2020-12, Vol.26 (12), p.2031-2043
• Main Authors: Immer, Carina, Hacker, Carolin, Wöhnert, Jens
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Biodegradation; Calorimetry; Conjugation; Gene regulation; Legionella pneumophila; Legionella pneumophila - genetics; Legionella pneumophila - metabolism; Magnetic resonance spectroscopy; NMR; Nuclear magnetic resonance; Nuclear Magnetic Resonance, Biomolecular - methods; Post-transcription; Protein Binding; Protein Domains; Proteins; Ribonucleic acid; RNA; RNA, Bacterial - chemistry; RNA, Bacterial - metabolism; RNA-binding protein; RNA-Binding Proteins - chemistry; RNA-Binding Proteins - metabolism; Structure-Activity Relationship; Titration; Uridine
• ISSN: 1355-8382; 1469-9001
• DOI: 10.1261/rna.077354.120

Small regulatory RNAs (sRNAs) play an important role for posttranscriptional gene regulation in bacteria. sRNAs recognize their target messenger RNAs (mRNAs) by base-pairing, which is often facilitated by interactions with the bacterial RNA-binding proteins Hfq or ProQ. The FinO/ProQ RNA-binding protein domain was first discovered in the bacterial repressor of conjugation, FinO. Since then, the functional role of FinO/ProQ-like proteins in posttranscriptional gene regulation was extensively studied in particular in the enterobacteria and and a wide range of sRNA-targets was identified for these proteins. In addition, enterobacterial ProQ homologs also recognize and protect the 3'-ends of a number of mRNAs from exonucleolytic degradation. However, the RNA-binding properties of FinO/ProQ proteins with regard to the recognition of different RNA targets are not yet fully understood. Here, we present the solution NMR structure of the so far functionally uncharacterized ProQ homolog Lpp1663 from as a newly confirmed member and a minimal model system of the FinO/ProQ protein family. In addition, we characterize the RNA-binding preferences of Lpp1663 with high resolution NMR spectroscopy and isothermal titration calorimetry (ITC). Our results suggest a binding preference for single-stranded uridine-rich RNAs in the vicinity of stable stem-loop structures. According to chemical shift perturbation experiments, the single-stranded U-rich RNAs interact mainly with a conserved RNA-binding surface on the concave site of Lpp1663.