NasR, a novel RNA-binding protein, mediates nitrate-responsive transcription antitermination of the Klebsiella oxytoca m5al nasF operon leader in Vitro

Chai, Weihang; Stewart, Valley

doi:10.1006/jmbi.1998.2105

Cited by 46 publications

(46 citation statements)

References 36 publications

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“…The latter consists of a unique class of proteins that have RNA-binding antiterminator function (17). Two response regulators containing this domain have been characterized, AmiR and NasR (18)(19)(20). These proteins associate directly with the 5Ј UTR of the nascent transcript to prevent formation of an intrinsic terminator.…”

Section: Resultsmentioning

confidence: 99%

“…The eut-associated response regulator shares this arrangement (10). Two proteins containing AN-TAR domains have previously been shown to interact with target mRNA transcripts to prevent formation of a transcription termination signal (16)(17)(18)(19)(20). We present evidence that the eut-associated 2-component regulatory system also interfaces with RNAmediated genetic control strategies.…”

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confidence: 80%

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Multiple posttranscriptional regulatory mechanisms partner to control ethanolamine utilization in Enterococcus faecalis

Fox¹,

Ramesh

Stearns

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

142

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Ethanolamine, a product of the breakdown of phosphatidylethanolamine from cell membranes, is abundant in the human intestinal tract and in processed foods. Effective utilization of ethanolamine as a carbon and nitrogen source may provide a survival advantage to bacteria that inhabit the gastrointestinal tract and may influence the virulence of pathogens. In this work, we describe a unique series of posttranscriptional regulatory strategies that influence expression of ethanolamine utilization genes (eut) in Enterococcus, Clostridium, and Listeria species. One of these mechanisms requires an unusual 2-component regulatory system. Regulation involves specific sensing of ethanolamine by a sensor histidine kinase (EutW), resulting in autophosphorylation and subsequent phosphoryl transfer to a response regulator (EutV) containing a RNA-binding domain. Our data suggests that EutV is likely to affect downstream gene expression by interacting with conserved transcription termination signals located within the eut locus. Breakdown of ethanolamine requires adenosylcobalamin (AdoCbl) as a cofactor, and, intriguingly, we also identify an intercistronic AdoCbl riboswitch that has a predicted structure different from previously established AdoCbl riboswitches. We demonstrate that association of AdoCbl to this riboswitch prevents formation of an intrinsic transcription terminator element located within the intercistronic region. Together, these results suggest an intricate and carefully coordinated interplay of multiple regulatory strategies for control of ethanolamine utilization genes. Gene expression appears to be directed by overlapping posttranscriptional regulatory mechanisms, each responding to a particular metabolic signal, conceptually akin to regulation by multiple DNAbinding transcription factors.riboswitch ͉ 2-component system

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 80%

Multiple posttranscriptional regulatory mechanisms partner to control ethanolamine utilization in Enterococcus faecalis

Fox¹,

Ramesh

Stearns

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

142

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“…For this purpose, the yield was rather low, so protein fusion methods were investigated as an alternative approach to Fur expression and purification. In the first instance, the maltose-binding protein (MBP) was chosen for fusion to Fur, since MBP has previously been used successfully for making fusions to transcriptional regulators (Chai & Stewart, 1998 ;Li et al, 1994). The fur gene was fused to the malE gene encoding MBP in the vector pMALc2.…”

Section: Resultsmentioning

confidence: 99%

The ferric uptake regulator of Pseudomonas aeruginosa has no essential cysteine residues and does not contain a structural zinc ion

et al. 2002

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The ferric uptake regulator (Fur) of Pseudomonas aeruginosa was expressed in Escherichia coli in its native form and as a fusion to the maltose-binding protein (MBP). Fur from the MBP fusion bound to MBP after proteolytic cleavage, and the two could only be separated by partial unfolding. The refolded protein was in the same conformation as native protein (as judged by circular dichroism and fluorescence spectroscopies) and was fully active in DNA-binding assays. As-prepared native Fur contained small amounts of Zn 2M that were easily removed by treatment with EDTA, and apo-protein could be reconstituted with approximately one Zn 2M ion per monomer. Thus, the P. aeruginosa Fur can probably accommodate a single Zn 2M ion bound to the metal-sensing site. The single cysteine residue of P. aeruginosa Fur aligns with a cysteine in other members of the Fur family that is essential for activity of the E. coli protein, and is believed to provide one of the ligands to a structural Zn 2M ion. This cysteine residue was shown to be dispensable for the in vivo activity of P. aeruginosa Fur, which is consistent with the suggestion that the P. aeruginosa protein does not contain a structural Zn 2M ion. Members of the Fur family contain a highly conserved His-His-Asp-His motif. Alanine substitutions of residues in this motif showed His-87 and His-89 of P. aeruginosa Fur to be essential for activity, whilst His-86 and Asp-88 are partially dispensable.

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“…We previously established that EA acts as the signal ligand that induces autophosphorylation of the histidine kinase of TCS-17 (HK17, aka EF1632 and EutW) that in turn phosphorylates the response regulator/transcription factor, RR17 (aka EF1633 and EutV) (11,15). The C-terminal domain of RR17 contains a putative ANTAR domain that generally regulates gene expression by an antitermination mechanism (8,11,34).…”

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confidence: 99%

Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B ₁₂ Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis

Baker

Perego

2011

J Bacteriol

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NasR, a novel RNA-binding protein, mediates nitrate-responsive transcription antitermination of the Klebsiella oxytoca m5al nasF operon leader in Vitro

Cited by 46 publications

References 36 publications

Multiple posttranscriptional regulatory mechanisms partner to control ethanolamine utilization in Enterococcus faecalis

Multiple posttranscriptional regulatory mechanisms partner to control ethanolamine utilization in Enterococcus faecalis

The ferric uptake regulator of Pseudomonas aeruginosa has no essential cysteine residues and does not contain a structural zinc ion

Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B ₁₂ Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis

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NasR, a novel RNA-binding protein, mediates nitrate-responsive transcription antitermination of the Klebsiella oxytoca m5al nasF operon leader in Vitro

Cited by 46 publications

References 36 publications

Multiple posttranscriptional regulatory mechanisms partner to control ethanolamine utilization in Enterococcus faecalis

Multiple posttranscriptional regulatory mechanisms partner to control ethanolamine utilization in Enterococcus faecalis

The ferric uptake regulator of Pseudomonas aeruginosa has no essential cysteine residues and does not contain a structural zinc ion

Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B 12 Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis

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Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B ₁₂ Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis