Identification and characterisation of proteins involved in riboregulation in Helicobacter pylori

Compared to other Gram-negative bacteria, the human pathogen Helicobacter pylori possess surprisingly few dedicated transcriptional regulators which raises the question how this organism coordinates complex cellular processes. We recently analysed the primary transcriptome of this organism to discover an unexpectedly large number of candidate regulatory RNAs (1), and now we are aiming to identify potential auxiliary protein factors.

Based on our previous studies in Salmonella (2, 3) we developed molecular methods to isolate in vivo and in vitro assembled RNA-protein complexes from H. pylori: (i) an RNA-based affinity chromatography strategy to identify protein binding partners of a given small RNA and (ii) a fast and easy method for the FLAG-tagging of proteins in H. pylori to facilitate co-immunoprecipitation of associated RNAs.

Using these methods, we have isolated functional RNA-protein complexes whose further characterisation might allow the discovery of new regulatory mechanisms that are involved in gene expression in this pathogen.