Expression of an inducible enzyme II fructose and activation of a cryptic enzyme II glucose in glucose-grown cells of spontaneous mutants of Streptococcus salivarius lacking the low-molecular-mass form of IIIman, a component of the phosphoenolpyruvate:mannose phosphotransferase system.
We have reported previously that the phosphoenolpyruvate:mannose phosphotransferase system (mannose PTS) of Streptococcus salivarius, consisting of an Enzyme II mannose (EIIman) and two forms of Enzyme III mannose (IIIman) with Mr values of 38,900 and 35,200, respectively, concomitantly transports and phosphorylates mannose, as well as glucose and fructose. In this paper, we report the presence, in S. salivarius, of alternative specific fructose and glucose PTSs encoded by inducible and cryptic genes, respectively. Protein phosphorylation experiments conducted with [32P]phosphoenolpyruvate have allowed us to identify by SDS-PAGE and autoradiography the EII fructose (EIIfru) (Mr 57,500) and the EII glucose (EIIglc) (Mr 58,700). No proteins corresponding to IIIfru or IIIglc could be detected. EIIfru phosphorylated fructose on the C-1 position rather than, as with the constitutive mannose PTS, on the C-6 position. Growth on fructose resulted in the induction of EIIfru as well as an increase of 1-phosphofructokinase activity. Nevertheless, the genes encoding these proteins were independently regulated. Studies carried out with spontaneous mutants lacking the low-molecular-mass form of IIIman (mutants A37, G29 and B31) showed that EIIfru was expressed in glucose-grown cells of strains G29 and B31, but not in strain A37, whereas the cryptic gene encoding EIIglc was activated in all three mutant strains. The results obtained with the mutants suggest that the three spontaneous mutants were not all mutated on the gene encoding IIIman although all of them lacked IIIman.
Bourassa
S
S
Department of Biochemistry (Sciences), Université Laval, Québec, Canada.
Vadeboncoeur
C
C
D004790:Enzyme Induction; D005632:Fructose / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D008358:Mannose / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D008970:Molecular Weight; D009154:Mutation; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D010732:Phosphofructokinase-1 / Q000378:metabolism; D013291:Streptococcus / Q000201:enzymology* / Q000235:genetics
D005632:Fructose; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C031629:phosphoenolpyruvate-glucose phosphotransferase; C037992:phosphoenolpyruvate-mannose phosphotransferase; D010732:Phosphofructokinase-1; C068652:1-phosphofructokinase; D005947:Glucose; D008358:Mannose