The phosphotransferase system of Streptomyces coelicolor.
We have investigated the crr gene of Streptomyces coelicolor that encodes a homologue of enzyme IIAGlucose of Escherichia coli, which, as a component of the phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS) plays a key role in carbon regulation by triggering glucose transport, carbon catabolite repression, and inducer exclusion. As in E. coli, the crr gene of S. coelicolor is genetically associated with the ptsI gene that encodes the general phosphotransferase enzyme I. The gene product IIACrr was overproduced, purified, and polyclonal antibodies were obtained. Western blot analysis revealed that IIACrr is expressed in vivo. The functionality of IIACrr was demonstrated by phosphoenolpyruvate-dependent phosphorylation via enzyme I and the histidine-containing phosphoryl carrier protein HPr. Phosphorylation was abolished when His72, which corresponds to the catalytic histidine of E. coli IIAGlucose, was mutated. The capacity of IIACrr to operate in sugar transport was shown by complementation of the E. coli glucose-PTS. The striking functional resemblance between IIACrr and IIAGlucose was further demonstrated by its ability to confer inducer exclusion of maltose to E. coli. A specific interaction of IIACrr with the maltose permease subunit MalK from Salmonella typhimurium was uncovered by surface plasmon resonance. These data suggest that this IIAGlucose-like protein may be involved in carbon metabolism in S. coelicolor.
Kamionka
Annette
A
Friedrich-Alexander-Universität Erlangen-Nürnberg, Lehrstuhl für Mikrobiologie, Erlangen, Germany.
Parche
Stephan
S
Nothaft
Harald
H
Siepelmeyer
Jörg
J
Jahreis
Knut
K
Titgemeyer
Fritz
F
D000595:Amino Acid Sequence; D001692:Biological Transport; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics / Q000378:metabolism*; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000201:enzymology*
D029968:Escherichia coli Proteins; C032242:crr protein, E coli; D010770:Phosphotransferases; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System