Dual posttranscriptional regulation via a cofactor-responsive mRNA leader

Laura M. Patterson-Fortin, Christopher A. Vakulskas, Elena Borisovna Yakhnina, Paul Lee Babitzke, Tony Romeo

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Abstract Riboswitches are cis-acting mRNA elements that regulate gene expression in response to ligand binding. Recently, a class of riboswitches was proposed to respond to the molybdenum cofactor (Moco), which serves as a redox center for metabolic enzymes. The 5′ leader of the Escherichia coli moaABCDE transcript exemplifies this candidate riboswitch class. This mRNA encodes enzymes for Moco biosynthesis, and moaA expression is feedback inhibited by Moco. Previous RNA-seq analyses showed that moaA mRNA copurified with the RNA binding protein CsrA (carbon storage regulator), suggesting that CsrA binds to this RNA in vivo. Among its global regulatory roles, CsrA represses stationary phase metabolism and activates central carbon metabolism. Here, we used gel mobility shift analysis to determine that CsrA binds specifically and with high affinity to the moaA 5′ mRNA leader. Northern blotting and studies with a series of chromosomal lacZ reporter fusions showed that CsrA posttranscriptionally activates moaA expression without altering moaA mRNA levels, indicative of translation control. Deletion analyses, nucleotide replacement studies and footprinting with CsrA-FeBABE identified two sites for CsrA binding. Toeprinting assays suggested that CsrA binding causes changes in moaA RNA structure. We propose that the moaA mRNA leader forms an aptamer, which serves as a target of posttranscriptional regulation by at least two different factors, Moco and the protein CsrA. While we are not aware of similar dual posttranscriptional regulatory mechanisms, additional examples are likely to emerge.

Original languageEnglish (US)
Pages (from-to)3662-3677
Number of pages16
JournalJournal of Molecular Biology
Volume425
Issue number19
DOIs
StatePublished - Oct 9 2013

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Carbon
Messenger RNA
Riboswitch
RNA
RNA-Binding Proteins
Coenzymes
Electrophoretic Mobility Shift Assay
Northern Blotting
Oxidation-Reduction
Nucleotides
Escherichia coli
Ligands
Gene Expression
molybdenum cofactor
Enzymes
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology

Cite this

Patterson-Fortin, Laura M. ; Vakulskas, Christopher A. ; Yakhnina, Elena Borisovna ; Babitzke, Paul Lee ; Romeo, Tony. / Dual posttranscriptional regulation via a cofactor-responsive mRNA leader. In: Journal of Molecular Biology. 2013 ; Vol. 425, No. 19. pp. 3662-3677.
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Dual posttranscriptional regulation via a cofactor-responsive mRNA leader. / Patterson-Fortin, Laura M.; Vakulskas, Christopher A.; Yakhnina, Elena Borisovna; Babitzke, Paul Lee; Romeo, Tony.

In: Journal of Molecular Biology, Vol. 425, No. 19, 09.10.2013, p. 3662-3677.

Research output: Contribution to journalArticle

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