Determinants of RNA recognition by the FinO domain of the Escherichia coli ProQ protein
| Autor: | Ewa M Stein, Mikołaj Olejniczak, Maciej M Basczok, Katherine E. Berry, Joanna Kwiatkowska, Chandra M Gravel |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Untranslated region
AcademicSubjects/SCI00010 Base pair Mutant RNA-binding protein Plasma protein binding Host Factor 1 Protein medicine.disease_cause Genetics Escherichia coli RNA and RNA-protein complexes medicine Binding site Nucleotide Motifs Hfq protein Regulation of gene expression Binding Sites biology Chemistry Escherichia coli Proteins RNA-Binding Proteins RNA Biochemistry Mutation biology.protein Terminator (franchise) Protein Binding |
| Zdroj: | Nucleic Acids Research |
| DOI: | 10.1101/2020.05.04.075150 |
| Popis: | The regulation of gene expression by small RNAs inEscherichia colidepends on RNA binding proteins Hfq and ProQ, which bind mostly distinct RNA pools. To understand how ProQ discriminates between RNA substrates, we compared its binding to six different RNA molecules. Full-length ProQ bound all six RNAs similarly, while the isolated N-terminal FinO domain (NTD) of ProQ specifically recognized RNAs with Rho-independent terminators. Analysis ofmalM3’-UTR mutants showed that tight RNA binding by the ProQ NTD required a terminator hairpin of at least two base pairs preceding an 3’ oligoU tail of at least four uridine residues. Substitution of an A-rich sequence on the 5’ side of the terminator to uridines strengthened the binding of several ProQ-specific RNAs to the Hfq protein, but not to the ProQ NTD. Substitution of the motif in themalM-3’ andcspE-3’ RNAs also conferred the ability to bind Hfq inE. colicells, as measured using a three-hybrid assay. In summary, these data suggest that the ProQ NTD specifically recognizes 3’ intrinsic terminators of RNA substrates, and that the discrimination between RNA ligands byE. coliProQ and Hfq depends both on positive determinants for binding to ProQ and negative determinants against binding to Hfq. |
| Databáze: | OpenAIRE |
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