Biochemical and genetic dissection of the RNA-binding surface of the FinO domain of Escherichia coli ProQ.

ABSTRACT: RNA-binding proteins play important roles in bacterial gene regulation through interactions with both coding and non-coding RNAs. ProQ is a FinO-domain protein that binds a large set of RNAs in Escherichia coli , though the details of how ProQ binds these RNAs remain unclear. In this study, we used a combination of in vivo and in vitro binding assays to confirm key structural features of E. coli ProQ's FinO domain and explore its mechanism of RNA interactions. Using a bacterial three-hybrid assay, we performed forward genetic screens to confirm the importance of the concave face of ProQ in RNA binding. Using gel shift assays, we directly probed the contributions of ten amino acids on ProQ binding to seven RNA targets. Certain residues (R58, Y70, and R80) were fo

SUBMITTER: Stein EM 

PROVIDER: S-EPMC10168233 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature