Protein kinase‐dependent HPr/CcpA interaction links glycolytic activity to carbon catabolite repression in Gram‐positive bacteria

Deutscher, Josef; Küster, Elke; Bergstedt, U; Charrier, Véronique; Hillen, Wolfgang

doi:10.1111/j.1365-2958.1995.tb02280.x

Cited by 336 publications

(272 citation statements)

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“…A specific interaction between CcpA and P-Ser-HPr was indeed demonstrated by retarded elution of P-SerHPr from CcpA-carrying columns 78) and later by nuclear resonance measurements. 79) In 1995, a complex of CcpA with P-Ser-HPr was verified to recognize the cre of the gnt operon with high affinity in footprinting experiments.…”

Section: Continuedmentioning

confidence: 80%

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Fujita

2009

Bioscience, Biotechnology, and Biochemistry

257

284

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Section: Continuedmentioning

confidence: 80%

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Fujita

2009

Bioscience, Biotechnology, and Biochemistry

257

284

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“…Phosphorylation of Ser 46 prevents the interaction of HPr with enzyme I, sugar-specific IIA proteins, and possibly with other protein domains (Reizer et al, 1984Deutscher et al, 1995;Ye & Saier, 1995). Although the enzyme I-binding site on B. subtilis HPr has not yet been determined, it has recently been shown to be indistinguishable from the enzyme IIAg"-binding site on HPr in the analogous E. coli proteins (van Nuland et al, 1995).…”

Section: Implications For Hpr Functionmentioning

confidence: 99%

“…HPr(Ser-P) interacts with a remarkably wide variety of proteins to control their activities, whereas neither the unphosphorylated form of HPr nor HPr(His-P) bind to these target proteins (Deutscher et al, 1995;Ye & Saier, 1995). The precise molecular basis for this selective binding is unknown, but it undoubtedly reflects electrostatic and/or conformational changes that accompany phosphorylation of HPr at Ser 46.…”

mentioning

confidence: 99%

Phosphorylation of serine‐46 in HPr, a key regulatory protein in bacteria, results in stabilization of its solution structure

et al. 1995

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The serine-phosphorylated form of histidine-containing protein (HPr), a component of the phosphoenolpyruvate:sugar phosphotransferase system from Bacillus subtilis, has been characterized by NMR spectroscopy and solvent denaturation studies. The results indicate that phosphorylation of Ser 46, the N-cap of a-helix-B, does not cause a conformational change but rather stabilizes the helix. Amide proton exchange rates in helix-B are decreased and phosphorylation stabilizes the protein to solvent and thermal denaturation, with a AAG of 0.7-0.8 kcal mol". A mutant in which Ser 46 is replaced by aspartic acid shows a similar stabilization, indicating that an electrostatic interaction between the negatively charged groups and the helix macrodipole contributes significantly to the stabilization.

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“…The trans-acting protein CcpA, a member of the LacI-GalR family of repressors (14), mediates CCR by binding to the cis-active operator sequence cre (catabolite responsive element) (15). An interaction of P-Ser-HPr with CcpA has been demonstrated in vitro, and this interaction is strengthened by FBP (16). Fujita et al (17) have shown that the protein complex formed between CcpA and P-Ser-HPr interacts specifically with the gnt cre.…”

mentioning

confidence: 99%

The Bacillus subtilis crh gene encodes a HPr-like protein involved in carbon catabolite repression

Galinier

Haiech

Kilhoffer

et al. 1997

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

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Carbon catabolite repression (CCR) of several Bacillus subtilis catabolic genes is mediated by ATPdependent phosphorylation of histidine-containing protein (HPr), a phosphocarrier protein of the phosphoenolpyruvate (PEP): sugar phosphotransferase system. In this study, we report the discovery of a new B. subtilis gene encoding a HPr-like protein, Crh (for catabolite repression HPr), composed of 85 amino acids. Crh exhibits 45% sequence identity with HPr, but the active site His-15 of HPr is replaced with a glutamine in Crh. Crh is therefore not phosphorylated by PEP and enzyme I, but is phosphorylated by ATP and the HPr kinase in the presence of fructose-1,6-bisphosphate. We determined Ser-46 as the site of phosphorylation in Crh by carrying out mass spectrometry with peptides obtained by tryptic digestion or CNBr cleavage. In a B. subtilis ptsH1 mutant strain, synthesis of ␤-xylosidase, inositol dehydrogenase, and levanase was only partially relieved from CCR. Additional disruption of the crh gene caused almost complete relief from CCR. In a ptsH1 crh1 mutant, producing HPr and Crh in which Ser-46 is replaced with a nonphosphorylatable alanyl residue, expression of ␤-xylosidase was also completely relieved from glucose repression. These results suggest that CCR of certain catabolic operons requires, in addition to CcpA, ATP-dependent phosphorylation of Crh, and HPr at Ser-46.The bacterial phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS) catalyzes the transport and concomitant phosphorylation of carbohydrates via a protein phosphorylation chain including PEP-dependent phosphorylation of His-15 in histidine-containing protein (HPr) by enzyme I (EI). P-His-HPr phosphorylates the sugar-specific EIIAs. In Gram-positive bacteria, the PTS regulates also induction and carbon catabolite repression (CCR) of numerous catabolic genes (1). The central regulatory protein involved in these various functions is HPr. In Gram-positive bacteria, this small phosphoryl transfer protein can be phosphorylated at a regulatory serine (Ser-46) by ATP and the HPr kinase (2, 3), in addition to phosphorylation at the catalytic His-15 by PEP and EI (4, 5). PEP-dependent and ATP-dependent phosphorylation of HPr interfere with each other-i.e., P-His-HPr is a poor substrate for the HPr kinase and P-Ser-HPr is a poor substrate for EI (6, 7). ATP-dependent phosphorylation of HPr is stimulated by glycolytic intermediates such as fructose-1,6-bisphosphate (FBP) in Enterococcus faecalis (6) and in Streptococcus pyogenes (7). It has been reported that FBP is also implicated in CCR of the Bacillus subtilis gnt and iol operons (8, 9), and a potential role of phosphorylation of HPr at Ser-46 in CCR has therefore been investigated (10). The gnt operon contains the genes gntRKPZ encoding the repressor GntR, gluconate kinase, gluconate permease, and a gluconate-6-Pdehydrogenase (11), whereas the iol operon is composed of 10 genes encoding enzymes presumably implicated in inositol metabolism, including iolG encoding inositol dehydro...

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Protein kinase‐dependent HPr/CcpA interaction links glycolytic activity to carbon catabolite repression in Gram‐positive bacteria

Cited by 336 publications

References 24 publications

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Carbon Catabolite Control of the Metabolic Network inBacillus subtilis

Phosphorylation of serine‐46 in HPr, a key regulatory protein in bacteria, results in stabilization of its solution structure

The Bacillus subtilis crh gene encodes a HPr-like protein involved in carbon catabolite repression

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