title: A cis-regulatory antisense RNA represses translation in Vibrio cholerae through extensive complementarity and proximity to the target locus.
abstract: As with all facultative pathogens, Vibrio cholerae must optimize its cellular processes to adapt to different environments with varying carbon sources and to environmental stresses. More specifically, in order to metabolize mannitol, V. cholerae must regulate the synthesis of MtlA, a mannitol transporter protein produced exclusively in the presence of mannitol. We previously showed that a cis-acting small RNA (sRNA) expressed by V. cholerae, MtlS, appears to post-transcriptionally downregulate the expression of mtlA and is produced in the absence of mannitol. We hypothesized that since it is complementary to the 5' untranslated region (UTR) of mtlA mRNA, MtlS may affect synthesis of MtlA by forming an mtlA-MtlS complex that blocks translation of the mRNA through occlusion of its ribosome binding site. To test this hypothesis, we used in vitro translation assays in order to examine the role MtlS plays in mtlA regulation and found that MtlS is sufficient to suppress translation of transcripts harboring the 5' UTR of mtlA. However, in a cellular context, the 5' UTR of mtlA is not sufficient for targeted repression by endogenous MtlS; additional segments from the coding region of mtlA play a role in the ability of the sRNA to regulate translation of mtlA mRNA. Additionally, proximity of transcription sites between the sRNA and mRNA significantly affects the efficacy of MtlS.
authors.affiliation: a Department of Chemistry; Pomona College ; Claremont , CA USA.
authors.forename: Howard
authors.initials: H
authors.lastname: Chang
authors.forename: John Michael
authors.initials: JM
authors.lastname: Replogle
authors.forename: Naomi
authors.initials: N
authors.lastname: Vather
authors.forename: Maya
authors.initials: M
authors.lastname: Tsao-Wu
authors.forename: Ronak
authors.initials: R
authors.lastname: Mistry
authors.forename: Jane M
authors.initials: JM
authors.lastname: Liu
mesh_terms: D020121:5' Untranslated Regions; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D020029:Base Pairing; D015964:Gene Expression Regulation, Bacterial; D056426:Genetic Loci; D008353:Mannitol / Q000378:metabolism; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism; D014734:Vibrio cholerae / Q000235:genetics* / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; D001426:Bacterial Proteins; D009004:Monosaccharide Transport Proteins; D034741:RNA, Small Interfering; D008353:Mannitol
keywords: MtlA; Vibrio cholerae; antisense RNA; cis-acting RNA; mannitol; post-transcriptional regulation; regulatory RNA
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