Regulation of the activity of the <i>Bacillus subtilis</i> antiterminator LicT by multiple PEP‐dependent, enzyme I‐ and HPr‐catalysed phosphorylation

Lindner, Cordula; Galinier, Anne; Hecker, Michael; Deutscher, Josef

doi:10.1046/j.1365-2958.1999.01262.x

Cited by 74 publications

(108 citation statements)

References 53 publications

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“…The second histidine in PRD1 participates in negative control of LicT and LacT (20,25). In contrast, the histidine(s) in PRD2 were shown to be necessary for activity of the dually controlled proteins SacT, LicT, and LacT (6,20,25), and preferential phosphorylation of these residues by HPr has been demonstrated for LicT (19). Based on these observations, it has been proposed that these proteins are stimulated by HPr-dependent phosphorylation of the histidines in PRD2 and inhibited by EII-dependent phosphorylation of the first histidine in PRD1.…”

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confidence: 98%

Regulation of the Escherichia coli Antiterminator Protein BglG by Phosphorylation at Multiple Sites and Evidence for Transfer of Phosphoryl Groups between Monomers

Görke

2003

Journal of Biological Chemistry

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Activity of antiterminator protein BglG regulating the ␤-glucoside operon in Escherichia coli is controlled by the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) in a dual manner. It requires HPr phosphorylation to be active, whereas phosphorylation by the ␤-glucoside-specific transport protein EII Bgl inhibits its activity. BglG and its relatives carry two PTS regulation domains (PRD1 and PRD2), each containing two conserved histidines. For BglG, histidine 208 in PRD2 was reported to be the negative phosphorylation site. In contrast, other antiterminators of this family are negatively regulated by phosphorylation of the first histidine in PRD1, and presumably activated by phosphorylation of the histidines in PRD2. In this work, a screen for mutant BglG proteins that escape repression by EII Bgl yielded exchanges of nine residues within PRD1, including conserved histidines His-101 and His-160, and C-terminally truncated proteins. Genetic and phosphorylation analyses indicate that His-101 in PRD1 is phosphorylated by EII Bgl and that His-160 contributes to negative regulation. His-208 in PRD2 is essential for BglG activity, suggesting that it is phosphorylated by HPr. Surprisingly, phosphorylation by HPr is not fully abolished by exchanges of His-208. However, phosphorylation by HPr is inhibited by exchanges in PRD1 and the phosphorylation of these mutants is restored in the presence of wild-type BglG. These results suggest that the activating phosphoryl group is transiently donated from HPr to PRD1 and subsequently transferred to His-208 of a second BglG monomer. The active His-208-phosphorylated BglG dimer can subsequently be inhibited in its activity by EII Bgl -catalyzed phosphorylation at His-101.

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confidence: 98%

Regulation of the Escherichia coli Antiterminator Protein BglG by Phosphorylation at Multiple Sites and Evidence for Transfer of Phosphoryl Groups between Monomers

Görke

2003

Journal of Biological Chemistry

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“…However, despite the overall homology shown by PRDs and the conservation of potential phosphorylation sites, the position(s) of phosphorylation and the effects on regulator activity appear to be highly variable. For example, the antiterminators LicT and SacY of B. subtilis have each been shown to be phosphorylated on three sites by HPr (only two of which are equivalent) ; however, the role of phosphorylation at these sites is not yet fully understood, and while LicT is activated SacY is, unusually, negatively regulated by HPr-mediated phosphorylation (Tortosa et al, 1997 ;Lindner et al, 1999). With respect to the transcriptional regulators shown in Fig.…”

Section: Discussionmentioning

confidence: 99%

Molecular analysis of the mannitol operon of Clostridium acetobutylicum encoding a phosphotransferase system and a putative PTS-modulated regulator

2001

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“…LicT was shown to be phosphorylated in vitro in the presence of the phosphorylated general PTS proteins on three of the four conserved histidines, i.e. the second conserved histidine in PRD1 and the two conserved histidines in PRD2 (28). Genetic analyses suggested that the two histidyl residues in PRD2 are important for LicT activation (28).…”

Section: Fig 3 Spr Analysis Of the Interaction Between The Prd Domamentioning

confidence: 99%

“…the second conserved histidine in PRD1 and the two conserved histidines in PRD2 (28). Genetic analyses suggested that the two histidyl residues in PRD2 are important for LicT activation (28). In contrast, SacY from B. subtilis was shown to be active in the absence of functional enzyme I and HPr.…”

Section: Fig 3 Spr Analysis Of the Interaction Between The Prd Domamentioning

confidence: 99%

Interactions between the PTS Regulation domains of the BglG Transcriptional Antiterminator from Escherichia coli

Fux

Nussbaum-Shochat

Amster‐Choder

2003

Journal of Biological Chemistry

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The E. coli BglG protein inhibits transcription termination within the bgl operon in the presence of ␤-glucosides. BglG represents a family of transcriptional antiterminators that bind to RNA sequences, which partially overlap rho-independent terminators, and prevent termination by stabilizing an alternative structure of the transcript. The activity of BglG is determined by its dimeric state, which is modulated by reversible phosphorylation catalyzed by BglF, a PTS permease. Only the non-phosphorylated BglG dimer binds to RNA and allows read-through of transcription. BglG is composed of three domains: an RNA-binding domain followed by two domains, PRD1 and PRD2 (PTS regulation domains), which are similar in their sequence and folding. Based on the three-dimensional structure of dimeric LicT, a BglG homologue from Bacillus subtilis, the interactions within the dimer are PRD1-PRD1 and PRD2-PRD2. We have shown before that PRD2 mediates homodimerization very efficiently. Using genetic systems and in vitro techniques that assay and characterize protein-protein interactions, we show here that the PRD1 dimerizes very slowly, but once it does, the homodimers are stable. These results support our model that formation of BglG dimers initiates with PRD2 dimerization followed by zipping up of two BglG monomers to create the active RNA-binding domain. Moreover, our results demonstrate that PRD1 and PRD2 heterodimerize efficiently in vitro and in vivo. The affinity among the PRDs is in the following order: PRD2-PRD2 > PRD1-PRD2

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Regulation of the activity of the Bacillus subtilis antiterminator LicT by multiple PEP‐dependent, enzyme I‐ and HPr‐catalysed phosphorylation

Cited by 74 publications

References 53 publications

Regulation of the Escherichia coli Antiterminator Protein BglG by Phosphorylation at Multiple Sites and Evidence for Transfer of Phosphoryl Groups between Monomers

Regulation of the Escherichia coli Antiterminator Protein BglG by Phosphorylation at Multiple Sites and Evidence for Transfer of Phosphoryl Groups between Monomers

Molecular analysis of the mannitol operon of Clostridium acetobutylicum encoding a phosphotransferase system and a putative PTS-modulated regulator

Interactions between the PTS Regulation domains of the BglG Transcriptional Antiterminator from Escherichia coli

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