A genetic approach for finding small RNAs regulators of genes of interest identifies RybC as regulating the DpiA/DpiB two-component system
Article first published online: 17 MAR 2009
Journal compilation © 2009 Blackwell Publishing Ltd. No claim to original US government works
Volume 72, Issue 3, pages 551–565, May 2009
How to Cite
Mandin, P. and Gottesman, S. (2009), A genetic approach for finding small RNAs regulators of genes of interest identifies RybC as regulating the DpiA/DpiB two-component system. Molecular Microbiology, 72: 551–565. doi: 10.1111/j.1365-2958.2009.06665.x
- Issue published online: 21 APR 2009
- Article first published online: 17 MAR 2009
- Accepted 6 March, 2009.
In Escherichia coli, the largest class of small regulatory RNAs binds to the RNA chaperone Hfq and regulates the stability and/or translation of specific mRNAs. While recent studies have shown that some mRNAs could be subject to post-transcriptional regulation by sRNAs (e.g. mRNAs found by co-immunoprecipitation with Hfq), no method has yet been described to identify small RNAs that regulate them. We developed a method to easily make translational fusions of genes of interest to the lacZ reporter gene, under the control of a PBAD-inducible promoter. A multicopy plasmid library of the E. coli genome can then be used to screen for small RNAs that affect the activity of the fusion. This screening method was first applied to the dpiB gene from the dpiBA operon, which encodes a two-component signal transduction system involved in the SOS response to β-lactams. One small RNA, RybC, was found to negatively regulate the expression of dpiB. Using mutants in the dpiB–lacZ fusion and compensatory mutations in the RybC sRNA, we demonstrate that RybC directly base pairs with the dpiBA mRNA.