NusG prevents transcriptional invasion of H-NS-silenced genes


Autoři: Lionello Bossi aff001;  Mathilde Ratel aff001;  Camille Laurent aff001;  Patricia Kerboriou aff001;  Andrew Camilli aff002;  Eric Eveno aff003;  Marc Boudvillain aff003;  Nara Figueroa-Bossi aff001
Působiště autorů: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Sud, Université Paris-Saclay, France aff001;  Department of Molecular Biology and Microbiology, Tufts University, Boston, MA, United States of America aff002;  Centre de Biophysique Moléculaire, CNRS UPR4301, rue Charles Sadron, France aff003
Vyšlo v časopise: NusG prevents transcriptional invasion of H-NS-silenced genes. PLoS Genet 15(10): e32767. doi:10.1371/journal.pgen.1008425
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008425

Souhrn

Evolutionarily conserved NusG protein enhances bacterial RNA polymerase processivity but can also promote transcription termination by binding to, and stimulating the activity of, Rho factor. Rho terminates transcription upon anchoring to cytidine-rich motifs, the so-called Rho utilization sites (Rut) in nascent RNA. Both NusG and Rho have been implicated in the silencing of horizontally-acquired A/T-rich DNA by nucleoid structuring protein H-NS. However, the relative roles of the two proteins in H-NS-mediated gene silencing remain incompletely defined. In the present study, a Salmonella strain carrying the nusG gene under the control of an arabinose-inducible repressor was used to assess the genome-wide response to NusG depletion. Results from two complementary approaches, i) screening lacZ protein fusions generated by random transposition and ii) transcriptomic analysis, converged to show that loss of NusG causes massive upregulation of Salmonella pathogenicity islands (SPIs) and other H-NS-silenced loci. A similar, although not identical, SPI-upregulated profile was observed in a strain with a mutation in the rho gene, Rho K130Q. Surprisingly, Rho mutation Y80C, which affects Rho’s primary RNA binding domain, had either no effect or made H-NS-mediated silencing of SPIs even tighter. Thus, while corroborating the notion that bound H-NS can trigger Rho-dependent transcription termination in vivo, these data suggest that H-NS-elicited termination occurs entirely through a NusG-dependent pathway and is less dependent on Rut site binding by Rho. We provide evidence that through Rho recruitment, and possibly through other still unidentified mechanisms, NusG prevents pervasive transcripts from elongating into H-NS-silenced regions. Failure to perform this function causes the feedforward activation of the entire Salmonella virulence program. These findings provide further insight into NusG/Rho contribution in H-NS-mediated gene silencing and underscore the importance of this contribution for the proper functioning of a global regulatory response in growing bacteria. The complete set of transcriptomic data is freely available for viewing through a user-friendly genome browser interface.

Klíčová slova:

Cloning – DNA transcription – Genetic loci – Pathogenesis – Salmonella – Arabinose – Transcriptional termination – RNA polymerase


Zdroje

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