Specific interaction of the RNA-binding domain of the Bacillus subtilis transcriptional antiterminator GlcT with its RNA target, RAT 1 1Edited by M. Gottesman

Langbein, Ines; Bachem, Steffi; Stülke, Jörg

doi:10.1006/jmbi.1999.3176

Cited by 37 publications

(52 citation statements)

References 36 publications

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“…These proteins (SacY, SacT, LicT, and GlcT) positively control the genes involved in the metabolism of carbohydrates that are taken up by the PTS; SacY and T are involved in sucrose metabolism, LicT in the metabolism of oligo--glucoside and aryl--glucoside, and GlcT in glucose assimilation. SacY and T [140][141][142] are antiterminators of the sacB (encoding levansucrase)-levB (endolevanase)-yveA, sacX (IIBC SacX )-sacY, and sacP (IIBC SacP )-sacA (sucrose-6-P hydrolase)-ywdA operons, whereas LicT 27,143) is an antiterminator of the licS gene (endo-1,3-1,4--glucanase) and the bglP (IIBCA Bgl )-bglH (6-P--glucosidase)-yxiE operon, and GlcT 144,145) is that of the ptsG (IICBA Glc )-ptsH (HPr)-ptsI (EI) operon. In the presence of the cognate PTS-sugars that activate these antiterminators, they bind to a conserved motif called the ribonucleic antiterminator (RAT), which is present in the untranslated leader-mRNAs of their target genes.…”

Section: Catabolite Control Mediated By Ccpb Ccpc Ccpn and Cggrmentioning

confidence: 99%

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Fujita

2009

Bioscience, Biotechnology, and Biochemistry

257

284

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Section: Catabolite Control Mediated By Ccpb Ccpc Ccpn and Cggrmentioning

confidence: 99%

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Fujita

2009

Bioscience, Biotechnology, and Biochemistry

257

284

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“…For example, the levanase gene of B. subtilis requires the action of a sigma 54-like regulator that activates transcription when cells are growing with levans or low levels of fructose (22) Similarly, a variety of transcriptional antiterminator proteins with homology to BglG, the transcriptional antiterminator of the Escherichia coli bglGFB operon, are present in Bacillus (SacY, SacT, LicT, and GlcT), Lactobacillus casei (LacT), Staphylococcus aureus, Lactococcus lactis, and S. mutans (BglG) (9,10,12,14,17,19,31,32,37), where they have been shown to control the expression of carbohydrate catabolism by allowing transcription of various operons. The binding site of antiterminator proteins, a ribonucleic antiterminator (RAT) sequence, has been well characterized in B. subtilis (2,20). In many cases, the activity of these antiterminators is modified by both the general and sugar-specific components of the phosphotransferase system (PTS) (36).…”

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confidence: 99%

Analysis of cis- and trans- Acting Factors Involved in Regulation of the Streptococcus mutans Fructanase Gene ( fruA )

Wen

Burne

2002

J Bacteriol

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There are two primary levels of control of the expression of the fructanase gene (fruA) of Streptococcus mutans: induction by levan, inulin, or sucrose and repression in the presence of glucose and other readily metabolized sugars. The goals of this study were to assess the functionality of putative cis-acting regulatory elements and to begin to identify the trans-acting factors involved in induction and catabolite repression of fruA. The fruA promoter and its derivatives generated by deletions and/or site-directed mutagenesis were fused to a promoterless chloramphenicol acetyltransferase (CAT) gene as a reporter, and strains carrying the transcriptional fusions were then analyzed for CAT activities in response to growth on various carbon sources. A dyadic sequence, ATGACA(TC)TGTCAT, located at ؊72 to ؊59 relative to the transcription initiation site was shown to be essential for expression of fruA. Inactivation of the genes that encode fructose-specific enzymes II resulted in elevated expression from the fruA promoter, suggesting negative regulation of fruA expression by the fructose phosphotransferase system. Mutagenesis of a terminator-like structure located in the 165-base 5 untranslated region of the fruA mRNA or insertional inactivation of antiterminator genes revealed that antitermination was not a mechanism controlling induction or repression of fruA, although the untranslated leader mRNA may play a role in optimal expression of fructanase. Deletion or mutation of a consensus catabolite response element alleviated glucose repression of fruA, but interestingly, inactivation of the ccpA gene had no discernible effect on catabolite repression of fruA. Accumulating data suggest that expression of fruA is regulated by a mechanism that has several unique features that distinguish it from archetypical polysaccharide catabolic operons of other gram-positive bacteria.

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“…without the PRDs, it is able to bind RNA and to exert antitermination activity constitutively (5,16). As shown in this work, the two phosphorylation events have different consequences; HPr-dependent phosphorylation has a slight stimulating effect on GlcT activity, whereas EII Glc -dependent phosphorylation inactivates the protein.…”

Section: Discussionmentioning

confidence: 99%

“…To express and purify GlcT fused to a hexahistidine sequence at the N terminus, we used plasmid pGP124 (5). Mutant alleles of glcT were obtained by two-step PCR mutagenesis essentially as described previously (16).…”

Section: Methodsmentioning

confidence: 99%

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Control of the Bacillus subtilis Antiterminator Protein GlcT by Phosphorylation

Schmalisch

Bachem

Stülke

2003

Journal of Biological Chemistry

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Bacillus subtilis transports glucose by the phosphotransferase system (PTS). The genes for this system are encoded in the ptsGHI operon, which is induced by glucose and depends on a termination/antitermination mechanism involving a riboswitch and the RNA-binding antitermination protein GlcT. In the absence of glucose, GlcT is inactive, and a terminator is formed in the leader region of the ptsG mRNA. If glucose is present, GlcT can bind to its RNA target and prevent transcription termination. The GlcT protein is composed of three domains, an N-terminal RNA binding domain and two PTS regulation domains, PTS regulation domain (PRD) I and PRD-II. In this work, we demonstrate that GlcT can be phosphorylated by two PTS proteins, HPr and the glucose-specific enzyme II (EII Glc ). HPr-dependent phosphorylation occurs on PRD-II and has a slight stimulatory effect on GlcT activity. In contrast, EII Glc phosphorylates the PRD-I of GlcT, and this phosphorylation inactivates GlcT. This latter phosphorylation event links the availability of glucose to the expression of the ptsGHI operon via the phosphorylation state of EII Glc and GlcT. This is the first in vitro demonstration of a direct phosphorylation of an antiterminator of the BglG family by the corresponding PTS permease.Glucose is the preferred source of carbon and energy for many bacteria. The sugar is transported into the cell and phosphorylated. The resulting glucose 6-phosphate can immediately feed into glycolysis. In Escherichia coli, Bacillus subtilis, and several other bacteria, glucose is taken up and concomitantly phosphorylated by the phosphoenolpyruvate:sugar phosphotransferase system (PTS).1 This system is made up of two general energy-coupling proteins, Enzyme I (EI) and HPr, and several multi-domain sugar specific permeases (Enzyme II, EII), which may exist as individual proteins or fused in a single polypeptide. In E. coli, the glucose-specific EII is composed of a membrane-bound protein comprising the actual transporter domain EIIC and the phosphotransfer domain EIIB. In addition, the EIIA domain is present as a cytoplasmic protein. In B. subtilis, all domains of the glucose permease are fused to form a single polypeptide with the domain arrangement EIICBA (1, 2).It was long considered that the genes encoding the components of the glucose PTS are constitutively expressed in bacteria. Although this is the case for the genes encoding the general proteins, ptsI and ptsH, the gene encoding EII Glc , ptsG, is induced by glucose in both E. coli and B. subtilis (1-3). In E. coli, regulation of ptsG expression is accomplished by the transcriptional repressor Mlc. In the presence of glucose, this repressor is sequestered by the non-phosphorylated EIICB Glc and, thus, is unable to repress ptsG transcription (3). In B. subtilis, glucose induction of ptsG expression is mediated by transcriptional antitermination. In the absence of glucose, transcription initiated at the ptsG promoter is terminated in the leader region of the mRNA. If glucose is present, an antiter...

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Specific interaction of the RNA-binding domain of the Bacillus subtilis transcriptional antiterminator GlcT with its RNA target, RAT 1 1Edited by M. Gottesman

Cited by 37 publications

References 36 publications

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Analysis of cis- and trans- Acting Factors Involved in Regulation of the Streptococcus mutans Fructanase Gene ( fruA )

Control of the Bacillus subtilis Antiterminator Protein GlcT by Phosphorylation

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