title: The SAM-responsive S(MK) box is a reversible riboswitch.
abstract: The S(MK) (SAM-III) box is an S-adenosylmethionine (SAM)-responsive riboswitch found in the 5' untranslated region of metK genes, encoding SAM synthetase, in many members of the Lactobacillales. SAM binding causes a structural rearrangement in the RNA that sequesters the Shine-Dalgarno (SD) sequence by pairing with a complementary anti-SD (ASD) sequence; sequestration of the SD sequence inhibits binding of the 30S ribosomal subunit and prevents translation initiation. We observed a slight increase in the half-life of the metK transcript in vivo when Enterococcus faecalis cells were depleted for SAM, but no significant change in overall transcript abundance, consistent with the model that this riboswitch regulates at the level of translation initiation. The half-life of the SAM-S(MK) box RNA complex in vitro is shorter than that of the metK transcript in vivo, raising the possibility of reversible binding of SAM. We used a fluorescence assay to directly visualize reversible switching between the SAM-free and SAM-bound conformations. We propose that the S(MK) box riboswitch can make multiple SAM-dependent regulatory decisions during the lifetime of the transcript in vivo, acting as a reversible switch that allows the cell to respond rapidly to fluctuations in SAM pools by modulating expression of the SAM synthetase gene.
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Angela M
authors.initials: AM
authors.lastname: Smith
authors.forename: Ryan T
authors.initials: RT
authors.lastname: Fuchs
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Grundy
authors.forename: Tina M
authors.initials: TM
authors.lastname: Henkin
mesh_terms: D020121:5' Untranslated Regions; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013293:Enterococcus faecalis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D008716:Methionine Adenosyltransferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D020871:RNA Stability; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020218:Response Elements; D058928:Riboswitch; D012436:S-Adenosylmethionine / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; D001426:Bacterial Proteins; D012329:RNA, Bacterial; D058928:Riboswitch; D012436:S-Adenosylmethionine; D008716:Methionine Adenosyltransferase