Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ
| Autor: | Smriti Pandey, Clara Wang, Oliver M Stockert, Hannah LeBlanc, Katherine E. Berry, Courtney L Hegner, Chandra M Gravel |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Untranslated region medicine.medical_specialty AcademicSubjects/SCI00010 Escherichia coli Proteins Protein domain RNA-Binding Proteins RNA Plasma protein binding Computational biology Biology medicine.disease_cause RNA Bacterial Genetic Techniques Protein Domains Molecular genetics RNA chaperone RNA and RNA-protein complexes Genetics medicine Escherichia coli Heat-Shock Proteins Protein Binding Genetic screen |
| Zdroj: | Nucleic Acids Research |
| Popis: | The FinO-domain-protein ProQ is an RNA-binding protein that has been known to play a role in osmoregulation in proteobacteria. Recently, ProQ has been shown to act as a global RNA-binding protein in Salmonella and E. coli, binding to dozens of small RNAs (sRNAs) and messenger RNAs (mRNAs) to regulate mRNA-expression levels through interactions with both 5′ and 3′ untranslated regions (UTRs). Despite excitement around ProQ as a novel global RNA-binding protein interacting with many sRNAs and mRNAs, and its potential to serve as a matchmaking RNA chaperone, significant gaps remain in our understanding of the molecular mechanisms ProQ uses to interact with RNA. In order to apply the tools of molecular genetics to this question, we have adapted a bacterial three-hybrid (B3H) assay to detect ProQ's interactions with target RNAs. Using domain truncations, site-directed mutagenesis and an unbiased forward genetic screen, we have identified a group of highly conserved residues on ProQ's NTD as the primary face for in vivo recognition of two RNAs, and propose that the NTD structure serves as an electrostatic scaffold to recognize the shape of an A-form RNA duplex. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|