Discovery of a ptsHI operon, which includes a third gene (ptsT), in the thermophile Bacillus stearothermophilus.
The discovery of ptsHI operon in Bacillus stearothermophilus XL-65-6 coupled with our previous report of a cel operon (Lai & Ingram, J Bacteriol 175, 6441-6450, 1993) demonstrates that this thermophilic organism contains all of the genes required for cellobiose uptake by the phosphoenolpyruvate-dependent phosphotransferase system (PTS). Genes encoding the two general PTS proteins, HPr (ptsH) and enzyme I (ptsI), were cloned and sequenced. These form an operon which includes a third small gene (ptsT) of unknown function (encoded product M(r) 18428). Both ptsH and ptsI were expressed at high levels from a single plasmid in Escherichia coli and complemented corresponding host mutations. Although the translated sequences for these genes were similar to homologues from Gram-positive mesophiles (64-77% identity), the B. stearothermophilus gene products were unusual in having a higher predicted pI and fewer negatively charged amino acid residues. Enzyme I also contained more alanine and leucine than mesophilic counterparts. Interestingly, ptsT inhibited the growth of E. coli ptsI mutants at 37 degrees C. No such inhibition was observed during incubation at a lower temperature (30 degrees C) or in E. coli DH5 alpha, which is wild-type for ptsI. The predicted translation product from ptsT contained a high proportion of basic amino acids (27%) and had a high predicted pI (pH 11.7), properties similar to bacterial histone-like proteins, but did not exhibit homology to any sequences in the current database. Regions upstream and downstream from the ptsHI operon contain genes with homology to Bacillus subtilis ptsG and wapA (wall-associated protein), respectively.
Lai
Xiaokuang
X
Department of Microbiology and Cell Science, IFAS, University of Florida, Gainesville, FL 32611, USA.
Ingram
Lonnie O
LO
Department of Microbiology and Cell Science, IFAS, University of Florida, Gainesville, FL 32611, USA.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D004926:Escherichia coli; D016368:Frameshift Mutation; D005798:Genes, Bacterial; D001411:Geobacillus stearothermophilus / Q000235:genetics*; D008969:Molecular Sequence Data; D009876:Operon; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000096:biosynthesis / Q000235:genetics*; D017852:Phosphotransferases (Nitrogenous Group Acceptor) / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013696:Temperature
D001426:Bacterial Proteins; C095036:PTST protein, Bacillus stearothermophilus; D011993:Recombinant Fusion Proteins; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C030182:phosphocarrier protein HPr; D017852:Phosphotransferases (Nitrogenous Group Acceptor); C022360:phosphoenolpyruvate-protein phosphotransferase