Novel phosphotransferase system genes revealed by bacterial genome analysis - a gene cluster encoding a unique Enzyme I and the proteins of a fructose-like permease system

Reizer, Jonathan; Reizer, Aiala; Saier, Milton H.

doi:10.1099/13500872-141-4-961

Cited by 43 publications

(32 citation statements)

References 53 publications

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“…As we show here, the gene talC of E. coli, also encodes a fructose-6-phosphate aldolase and not a transaldolase as proposed earlier (19). The gene product shows a high degree of similarity with FSA.…”

Section: Discussionmentioning

confidence: 56%

“…We have cloned an open reading frame from the E. coli chromosome that had been assumed earlier to be a transaldolase or a transaldolase-related protein (19). 2 Here we show that this gene encodes a novel enzyme activity, fructose-6-phosphate aldolase.…”

Section: Discussionmentioning

confidence: 99%

“…As a side reaction, formation of fructose-6-P from dihydroxyacetone and glyceraldehyde-3-P is known, but the corresponding aldol cleavage reaction has not been documented (3). Recently, a group of gene sequences presumably encoding transaldolase-like proteins (19) has been reported as outcome of total genome analyses of various Eubacteria and Archaebacteria. We have cloned two of these sequences (mipB and talC) from the genome of E. coli K-12.…”

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confidence: 99%

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Fructose-6-phosphate Aldolase Is a Novel Class I Aldolase from Escherichia coli and Is Related to a Novel Group of Bacterial Transaldolases

Schürmann

Sprenger

2001

Journal of Biological Chemistry

193

171

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We have cloned an open reading frame from the Escherichia coli K-12 chromosome that had been assumed earlier to be a transaldolase or a transaldolase-related protein, termed MipB. Here we show that instead a novel enzyme activity, fructose-6-phosphate aldolase, is encoded by this open reading frame, which is the first report of an enzyme that catalyzes an aldol cleavage of fructose 6-phosphate from any organism. We propose the name FSA (for fructose-six phosphate aldolase; gene name fsa). The recombinant protein was purified to apparent homogeneity by anion exchange and gel permeation chromatography with a yield of 40 mg of protein from 1 liter of culture. By using electrospray tandem mass spectroscopy, a molecular weight of 22,998 per subunit was determined. From gel filtration a size of 257,000 (؎ 20,000) was calculated. The enzyme most likely forms either a decamer or dodecamer of identical subunits. The purified enzyme displayed a V max of 7 units mg ؊1 of protein for fructose 6-phosphate cleavage (at 30°C, pH 8.5 in 50 mM glycylglycine buffer). For the aldolization reaction a V max of 45 units mg ؊1 of protein was found; K m values for the substrates were 9 mM for fructose 6-phosphate, 35 mM for dihydroxyacetone, and 0.8 mM for glyceraldehyde 3-phosphate. FSA did not utilize fructose, fructose 1-phosphate, fructose 1,6-bisphosphate, or dihydroxyacetone phosphate. FSA is not inhibited by EDTA which points to a metal-independent mode of action. The lysine 85 residue is essential for its action as its exchange to arginine (K85R) resulted in complete loss of activity in line with the assumption that the reaction mechanism involves a Schiff base formation through this lysine residue (class I aldolase). Another fsa-related gene, talC of Escherichia coli, was shown to also encode fructose-6-phosphate aldolase activity and not a transaldolase as proposed earlier.

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Section: Discussionmentioning

confidence: 56%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Fructose-6-phosphate Aldolase Is a Novel Class I Aldolase from Escherichia coli and Is Related to a Novel Group of Bacterial Transaldolases

Schürmann

Sprenger

2001

Journal of Biological Chemistry

193

171

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“…1C). I Ntr is a multidomain protein consisting of a homologue of the PTS EI and a domain also found in NifA activator proteins (30).…”

Section: Proteobacteriamentioning

confidence: 99%

CcpA-Dependent Carbon Catabolite Repression in Bacteria

Warner

Lolkema

2003

Microbiol Mol Biol Rev

236

219

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Carbon catabolite repression (CCR) by transcriptional regulators follows different mechanisms in gram-positive and gram-negative bacteria. In gram-positive bacteria, CcpA-dependent CCR is mediated by phosphorylation of the phosphoenolpyruvate:sugar phosphotransferase system intermediate HPr at a serine residue at the expense of ATP. The reaction is catalyzed by HPr kinase, which is activated by glycolytic intermediates. In this review, the distribution of CcpA-dependent CCR among bacteria is investigated by searching the public databases for homologues of HPr kinase and HPr-like proteins throughout the bacterial kingdom and by analyzing their properties. Homologues of HPr kinase are commonly observed in the phylum Firmicutes but are also found in the phyla Proteobacteria, Fusobacteria, Spirochaetes, and Chlorobi, suggesting that CcpA-dependent CCR is not restricted to gram-positive bacteria. In the α and β subdivisions of the Proteobacteria, the presence of HPr kinase appears to be common, while in the γ subdivision it is more of an exception. The genes coding for the HPr kinase homologues of the Proteobacteria are in a gene cluster together with an HPr-like protein, termed XPr, suggesting a functional relationship. Moreover, the XPr proteins contain the serine phosphorylation sequence motif. Remarkably, the analysis suggests a possible relation between CcpA-dependent gene regulation and the nitrogen regulation system (Ntr) found in the γ subdivision of the Proteobacteria. The relation is suggested by the clustering of CCR and Ntr components on the genome of members of the Proteobacteria and by the close phylogenetic relationship between XPr and NPr, the HPr-like protein in the Ntr system. In bacteria in the phylum Proteobacteria that contain HPr kinase and XPr, the latter may be at the center of a complex regulatory network involving both CCR and the Ntr system

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“…The diversity of the fructose-like PTS proteins including HrsA noted above could result from domain shuffling and splicing which must have occurred during the evolution of these and other PTS proteins as previously described. 33 ) The truncated HrsA can work as a heat-responsible regulator of ompC expression in T02 (micF mutant of RUIOI2), but cannot in Kl108 (ompR mutant of RUIOI2) (Figs. I and 2).…”

Section: Discussionmentioning

confidence: 99%