The minimal ProQ protein of Neisseria meningitidis is a global RNA-binding protein that operates in parallel with Hfq [RNA-Seq]

In this study the RNA targetome of the RNA-binding protein ProQ was determined in Neisseria meninigitidis serogroup C strain 8013 by RNA-seq. Overall design: To uncover the RNA targetome of the RNA-binding protein ProQ of Neisseria meningitidis (N.meninigitidis), bacterial samples were collected of three biological replicates of N.meninigitidis strain 8013 wt and proQ deletion in which the CDS of ProQ was replaced by an antibiotics cassette. Bacterial cultures grown to OD600nm of 2.0 were snapfrozen in liquid nitrogen prior to total RNA-extraction with the hot phenol method and DNase digestion. cDNA libraries of RNA-seq-samples were constructed by Vertis Biotechnology AG, Munich, Germany. To deplete ribosomal transcripts, RNA-seq samples were treated with the Ribo-Zero “Bacteria” kit (Illumina) followed by RNA fragmentation and adapter ligation. cDNA libraries of RNA-seq samples were pooled on an Illumina NextSeq 500 high-output flow cell and sequenced in single-read mode (1x76 cycles).