PMID:8132321

Citation	Boyd, DA, Cvitkovitch, DG and Hamilton, IR (1994) Sequence and expression of the genes for HPr (ptsH) and enzyme I (ptsI) of the phosphoenolpyruvate-dependent phosphotransferase transport system from Streptococcus mutans. Infect. Immun. 62:1156-65
Abstract	We report the sequencing of a 2,242-bp region of the Streptococcus mutants NG5 genome containing the genes for ptsH and ptsI, which encode HPr and enzyme I (EI), respectively, of the phosphoenolpyruvate-dependent phosphotransferase transport system. The sequence was obtained from two cloned overlapping genomic fragments; one expresses HPr and a truncated EI, while the other expresses a full-length EI in Escherichia coli, as determined by Western immunoblotting. The ptsI gene appeared to be expressed from a region located in the ptsH gene. The S. mutans NG5 pts operon does not appear to be linked to other phosphotransferase transport system proteins as has been found in other bacteria. A positive fermentation pattern on MacConkey-glucose plates by an E. coli ptsI mutant harboring the S. mutans NG5 ptsI gene on a plasmid indicated that the S. mutans NG5 EI can complement a defect in the E. coli gene. This was confirmed by protein phosphorylation experiments with 32P-labeled phosphoenolpyruvate indicating phosphotransfer from the S. mutans NG5 EI to the E. coli HPr. Two forms of the cloned EI, both truncated to varying degrees in the C-terminal region, were inefficiently phosphorylated and unable to complement fully the ptsI defect in the E. coli mutant. The deduced amino acid sequence of HPr shows a high degree of homology, particularly around the active site, to the same protein from other gram-positive bacteria, notably, S. salivarius, and to a lesser extent with those of gram-negative bacteria. The deduced amino acid sequence of S. mutans NG5 EI also shares several regions of homology with other sequenced EIs, notably, with the region around the active site, a region that contains the only conserved cystidyl residue among the various proteins and which may be involved in substrate binding.
Links	PubMed PMC186246
Keywords	Amino Acid Sequence; Bacterial Proteins; Base Composition; Base Sequence; Chromosome Mapping; Cloning, Molecular; Escherichia coli/genetics; Genes, Bacterial; Molecular Sequence Data; Phosphoenolpyruvate Sugar Phosphotransferase System/chemistry; Phosphoenolpyruvate Sugar Phosphotransferase System/genetics; Phosphorylation; Phosphotransferases (Nitrogenous Group Acceptor)/chemistry; Phosphotransferases (Nitrogenous Group Acceptor)/genetics; Streptococcus mutans/genetics
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
STRMU:PT1		GO:0009401: phosphoenolpyruvate-dependent sugar phosphotransferase system	ECO:0000247:	UniProtKB:P08839	P		Fig. 3 nucleotide sequence of ptsI gene of S. mutans shows 47.7% identity with E1 of E. coli. E1 and HPr products of ptsI genes in S. mutans are also shown to be expressed in E. coli and E1 can complement a ptsI defect in E. coli.	complete CACAO 4955
STRMU:PT1	involved_in	GO:0009401: phosphoenolpyruvate-dependent sugar phosphotransferase system	ECO:0000247: sequence alignment evidence used in manual assertion	UniProtKB:P08839	P		Seeded From UniProt	complete
STACT:PT1		GO:0009401: phosphoenolpyruvate-dependent sugar phosphotransferase system	ECO:0000247:	UniProtKB:P08839	P		E1 protein alignments of S. carnosus and E. coli seen in Fig. 6B shows several regions of conserved regions and homology. E1 is a product of the ptsI gene in E. coli, which has IDA to PEP-dependent phosphotransferase activity.	complete
STRMU:PTHP		GO:0009401: phosphoenolpyruvate-dependent sugar phosphotransferase system	ECO:0000247:	UniProtKB:P0AA04	P		E1 and HPr are products of ptsI genes in S. mutans that are shown to be expressed in E. coli (which has IDA evidence of activity). Fig. 3 nucleotide sequence of ptsH gene of S. mutans shows 37.7% identity with HPr of E. coli. Fig. 6A also shows the HPr alignments of S. mutans and E. coli exhibiting high homology.	complete
STRSL:PTHP		GO:0009401: phosphoenolpyruvate-dependent sugar phosphotransferase system	ECO:0000247:	UniProtKB:P0AA04	P		HPr protein alignments of S. salivarius and E. coli seen in Fig. 6A exhbits high homology. HPr is a product of the ptsH gene in E. coli, which has IDA to PEP-dependent phosphotransferase system.	complete

References

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