| Autor: |
Morón Á; Department of Cell Biology, Facultad de Ciencias Biológicas, Universidad Complutense de Madrid, 28040 Madrid, Spain.; Laboratory of Intracellular Bacterial Pathogens, National Centre for Biotechnology (CNB)-CSIC, 28049 Madrid, Spain., Ortiz-Miravalles L; Laboratory of Intracellular Bacterial Pathogens, National Centre for Biotechnology (CNB)-CSIC, 28049 Madrid, Spain.; Department of Molecular Biology, Universidad Autónoma de Madrid, Centro de Biologia Molecular Severo Ochoa (CBM) CSIC-UAM, 28049 Madrid, Spain., Peñalver M; Laboratory of Intracellular Bacterial Pathogens, National Centre for Biotechnology (CNB)-CSIC, 28049 Madrid, Spain.; Department of Molecular Biology, Universidad Autónoma de Madrid, Centro de Biologia Molecular Severo Ochoa (CBM) CSIC-UAM, 28049 Madrid, Spain., García-Del Portillo F; Laboratory of Intracellular Bacterial Pathogens, National Centre for Biotechnology (CNB)-CSIC, 28049 Madrid, Spain., Pucciarelli MG; Laboratory of Intracellular Bacterial Pathogens, National Centre for Biotechnology (CNB)-CSIC, 28049 Madrid, Spain.; Department of Molecular Biology, Universidad Autónoma de Madrid, Centro de Biologia Molecular Severo Ochoa (CBM) CSIC-UAM, 28049 Madrid, Spain., Ortega AD; Department of Cell Biology, Facultad de Ciencias Biológicas, Universidad Complutense de Madrid, 28040 Madrid, Spain.; Department of Molecular Biology, Universidad Autónoma de Madrid, Centro de Biologia Molecular Severo Ochoa (CBM) CSIC-UAM, 28049 Madrid, Spain. |
| Abstrakt: |
Listeria pathogenicity island 1 (LIPI-1) is a genetic region containing a cluster of genes essential for virulence of the bacterial pathogen Listeria monocytogenes . Main virulence factors in LIPI-1 include long 5' untranslated regions (5'UTRs), among which is Rli51, a small RNA (sRNA) in the 5'UTR of the Zn-metalloprotease-coding mpl . So far, Rli51 function and molecular mechanisms have remained obscure. Here, we show that Rli51 exhibits a dual mechanism of regulation, functioning as a cis - and as a trans -acting sRNA. Under nutrient-rich conditions, rli51-mpl transcription is prematurely terminated, releasing a short 121-nucleotide-long sRNA. Rli51 is predicted to function as a transcription attenuator that can fold into either a terminator or a thermodynamically more stable antiterminator. We show that the sRNA Rli21/RliI binds to a single-stranded RNA loop in Rli51, which is essential to mediate premature transcription termination, suggesting that sRNA binding could stabilize the terminator fold. During intracellular infection, rli51 transcription is increased, which generates a higher abundance of the short Rli51 sRNA and allows for transcriptional read-through into mpl . Comparative intracellular bacterial transcriptomics in rli51 -null mutants and the wild-type reference strain EGD-e suggests that Rli51 upregulates iron-scavenging proteins and downregulates virulence factors from LIPI-1. MS2 affinity purification confirmed that Rli51 binds transcripts of the heme-binding protein Lmo2186 and Lmo0937 in vivo. These results prove that Rli51 functions as a trans -acting sRNA in intracellular bacteria. Our research shows a growth condition-dependent mechanism of regulation for Rli51, preventing unintended mpl transcription in extracellular bacteria and regulating genes important for virulence in intracellular bacteria. |
