Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system: purification and characterization of the mannitol-specific enzyme IIImtl of Staphylococcus aureus and Staphylococcus carnosus and homology with the enzyme IImtl of Escherichia coli

Reiche, B; Frank, Rainer; Deutscher, Josef; Meyer, N.; Hengstenberg, Wolfgang

doi:10.1021/bi00417a047

Cited by 46 publications

(29 citation statements)

References 17 publications

(9 reference statements)

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“…A comparison of the crystal structures of E. coli EIIA Ntr and the EIIA Mtl domain shows that the fold of both proteins as well as the structure around the phosphorylation site, His-73 in EIIA Ntr and His-65 in EIIA Mtl (703), are nearly identical (73,905 (717). These findings are consistent with a regulatory role of EIIA Ntr through binding rather than through phosphoryl group transfer.…”

Section: The Pts Ntrsupporting

confidence: 55%

How Phosphotransferase System-Related Protein Phosphorylation Regulates Carbohydrate Metabolism in Bacteria

Deutscher

Francke

Postma

2006

Microbiol Mol Biol Rev

1,193

769

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SUMMARY The phosphoenolpyruvate(PEP):carbohydrate phosphotransferase system (PTS) is found only in bacteria, where it catalyzes the transport and phosphorylation of numerous monosaccharides, disaccharides, amino sugars, polyols, and other sugar derivatives. To carry out its catalytic function in sugar transport and phosphorylation, the PTS uses PEP as an energy source and phosphoryl donor. The phosphoryl group of PEP is usually transferred via four distinct proteins (domains) to the transported sugar bound to the respective membrane component(s) (EIIC and EIID) of the PTS. The organization of the PTS as a four-step phosphoryl transfer system, in which all P derivatives exhibit similar energy (phosphorylation occurs at histidyl or cysteyl residues), is surprising, as a single protein (or domain) coupling energy transfer and sugar phosphorylation would be sufficient for PTS function. A possible explanation for the complexity of the PTS was provided by the discovery that the PTS also carries out numerous regulatory functions. Depending on their phosphorylation state, the four proteins (domains) forming the PTS phosphorylation cascade (EI, HPr, EIIA, and EIIB) can phosphorylate or interact with numerous non-PTS proteins and thereby regulate their activity. In addition, in certain bacteria, one of the PTS components (HPr) is phosphorylated by ATP at a seryl residue, which increases the complexity of PTS-mediated regulation. In this review, we try to summarize the known protein phosphorylation-related regulatory functions of the PTS. As we shall see, the PTS regulation network not only controls carbohydrate uptake and metabolism but also interferes with the utilization of nitrogen and phosphorus and the virulence of certain pathogens.

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Section: The Pts Ntrsupporting

confidence: 55%

How Phosphotransferase System-Related Protein Phosphorylation Regulates Carbohydrate Metabolism in Bacteria

Deutscher

Francke

Postma

2006

Microbiol Mol Biol Rev

1,193

769

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“…We have recently demonstrated that mannitol transport in S. mutans has an EIIA component (Honeyman & Curtiss, 1992). This is similar to the mannitol PTSs found in other Gram-positive organisms (Fischer et al, 1989(Fischer et al, , 1991Reiche et al, 1988 ;Henstra et al, 1996).…”

Section: Introductionsupporting

confidence: 83%

“…However, the second possibility seems remote because the only genes in any other mannitol PTS system which are necessary for mannitol utilization are mtlA, mtlF and mtlD (Fischer & Hengstenberg, 1992 ;Fischer et al, 1991 ;Henstra et al, 1996Henstra et al, , 1999Lee & Saier, 1983 ;Reiche et al, 1988). These genes have not been interrupted in this case, and the observed effects are therefore probably due to the polar insertion of the interposon.…”

Section: Insertional Inactivation Of Mtlrmentioning

confidence: 86%

“…The mannitol PTS in E. coli utilizes a single sugar-specific Enzyme IICBA molecule. In other organisms, mannitol transport may utilize an EIICB-EIIA pair as depicted for glucose transport in E. coli (Fischer et al, 1989(Fischer et al, , 1991Reiche et al, 1988). We have recently demonstrated that mannitol transport in S. mutans has an EIIA component (Honeyman & Curtiss, 1992).…”

Section: Introductionmentioning

confidence: 99%

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The mannitol-specific enzyme II (mtlA) gene and the mtlR gene of the PTS of Streptococcus mutans The GenBank accession number for the sequence reported in this paper is AF210133.

Honeyman

Curtiss

2000

Microbiology

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The phosphoenolpyruvate-dependent phosphotransferase system (PTS) is widely found among Gram-positive bacteria. It is the major source of carbohydrate transport in the dental pathogen Streptococcus mutans. The transported carbohydrates are fermented to produce large amounts of lactic acid which initiates dental caries. The authors have isolated the S. mutans gene for the mannitol-specific Enzyme II (EII) component of the PTS, mtlA, and the adjacent mtlR gene, which is located in the same operon. The mtlR gene is located between mtlA and the genes mtlF and mtlD. The nucleotide sequence of the mtlA and mtlR loci has been determined. The deduced mtlA gene product of S. mutans consists of 589 amino acids with a molecular mass of 620 kDa. It exhibits similarity with the mtlA gene products from other organisms. However, the similarity between these proteins is generally restricted to the 470 amino-terminal residues of the S. mutans protein. This region would correspond to the EIICB domains of the PTS. The authors have previously shown that the S. mutans mtlF gene product exhibits 766% similarity to the carboxyl-terminal 143 amino acids of the Escherichia coli mtlA product and that the mtlF gene encodes the EIIA domain of the PTS. Thus, the genes that encode the EIICB and the EIIA domains are separated by approximately 2250 bp. In many organisms, all of the EII domains may be fused together to form one molecule. The fact that these domains are separated by this distance in S. mutans supports the hypothesis that various functional domains of the PTS have been rearranged during evolution. The sequence of the 119 carboxyl-terminal amino acids of the S. mutans mtlA gene product also displays homology to the carboxyl-terminal end of the EIIB domain of various mannitol PTSs. Thus, this domain may have been duplicated in S. mutans during evolution of the operon. The mtlR gene is located in the same operon structure as mtlA but these loci are separated by an intragenic space. The precise 5' end of the mtlR locus cannot be determined either by in vitro transcription-translation assays or based upon nucleotide sequence analysis because of the apparent lack of a ribosome-binding site preceding the gene. The deduced mtlR gene product, which consists of approximately 650 amino acids with a molecular mass of 753 kDa, exhibits limited similarity to several potential transcriptional regulators. However, the exact function of this locus is currently unknown.

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“…Expression of the various enzymes was induced by growth in the presence of the appropriate carbon source. Gluconate kinase, inositol dehydrogenase, and glucitol dehydrogenase activities were determined as described previously (16,30); mannitol PTS activity was determined according to the method of Reiche et al (32); mannitol-1-P dehydrogenase activity was measured according to the method of Novotny et al (31); glycerol kinase activity was measured according to the method of Deutscher and Sauerwald (11); oa-glucosidase activity was determined as described previously (4) by using p-nitrophenyl-ao-D-glucopyranoside as substrate, and ,B-galactosidase activity was measured according to the method of Miller (27) (Table 2). Repression of these enzymes by fructose was also no longer observed (9).…”

Section: Methodsmentioning

confidence: 99%

Loss of protein kinase-catalyzed phosphorylation of HPr, a phosphocarrier protein of the phosphotransferase system, by mutation of the ptsH gene confers catabolite repression resistance to several catabolic genes of Bacillus subtilis

et al. 1994

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In gram-positive bacteria, HPr, a phosphocarrier protein of the phosphoenolpyruvate:sugar phosphotransferase system (PTS), is phosphorylated by an ATP-dependent, metabolite-activated protein kinase on seryl residue 46. In a Bacillus subtilis mutant strain in which Ser-46 of HPr was replaced with a nonphosphorylatable alanyl residue (ptsHl mutation), synthesis of gluconate kinase, glucitol dehydrogenase, mannitol-l-P dehydrogenase and the mannitol-specific PTS permease was completely relieved from repression by glucose, fructose, or mannitol, whereas synthesis of inositol dehydrogenase was partially relieved from catabolite repression and synthesis of oa-glucosidase and glycerol kinase was still subject to catabolite repression. When the S46A mutation in HPr was reverted to give S46 wild-type HPr, expression of gluconate kinase and glucitol dehydrogenase regained full sensitivity to repression by PTS sugars. These results suggest that phosphorylation of HPr at Ser-46 is directly or indirectly involved in catabolite repression. A strain deleted for the ptsGHI genes was transformed with plasmids expressing either the wild-type ptsH gene or various S46 mutant ptsH genes (S46A or S46D). Expression of the gene encoding S46D HPr, having a structure similar to that of P-ser-HPr according to nuclear magnetic resonance data, caused significant reduction of gluconate kinase activity, whereas expression of the genes encoding wild-type or S46A HPr had no effect on this enzyme activity. When the promoterless lacZ gene was put under the control of the gnt promoter and was subsequently incorporated into the amyE gene on the B. subtilis chromosome, expression of 1-galactosidase was inducible by gluconate and repressed by glucose. However, we observed no repression of I-galactosidase activity in a strain carrying the ptsHl mutation. Additionally, we investigated a ccpA mutant strain and observed that all of the enzymes which we found to be relieved from carbon catabolite repression in the ptsHl mutant strain were also insensitive to catabolite repression in the ccpA mutant. Enzymes that were repressed in the ptsHl mutant were also repressed in the ccpA mutant.Catabolite-repressible genes in the gram-positive bacterium Bacillus subtilis are controlled by multiple mechanisms rather than by a single global regulatory system. These mechanisms are known to differ from those operating in enteric bacteria such as Escherichia coli, but the molecular details are poorly understood (15,40,46

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Staphylococcal phosphoenolpyruvate-dependent phosphotransferase system: purification and characterization of the mannitol-specific enzyme IIImtl of Staphylococcus aureus and Staphylococcus carnosus and homology with the enzyme IImtl of Escherichia coli

Cited by 46 publications

References 17 publications

How Phosphotransferase System-Related Protein Phosphorylation Regulates Carbohydrate Metabolism in Bacteria

How Phosphotransferase System-Related Protein Phosphorylation Regulates Carbohydrate Metabolism in Bacteria

The mannitol-specific enzyme II (mtlA) gene and the mtlR gene of the PTS of Streptococcus mutans The GenBank accession number for the sequence reported in this paper is AF210133.

Loss of protein kinase-catalyzed phosphorylation of HPr, a phosphocarrier protein of the phosphotransferase system, by mutation of the ptsH gene confers catabolite repression resistance to several catabolic genes of Bacillus subtilis

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