sigma54-Mediated control of the mannose phosphotransferase sytem in Lactobacillus plantarum impacts on carbohydrate metabolism.
Sigma factors direct specific binding of the bacterial RNA polymerase to the promoter. Here we present the elucidation of the sigma(54 ) regulon in Lactobacillus plantarum. A sequence-based regulon prediction of sigma(54)-dependent promoters revealed an operon encoding a mannose phosphotransferase system (PTS) as the best candidate for sigma(54)-mediated control. A sigma (54) (rpoN) mutant derivative did not grow on mannose, confirming this prediction. Additional mutational analyses established the presence of one functional mannose PTS in L. plantarum, the expression of which is controlled by sigma(54) in concert with the sigma(54)-activator ManR. Genome-wide transcription comparison of the wild-type and the rpoN-deletion strain revealed nine upregulated genes in the wild-type, including the genes of the mannose PTS, and 21 upregulated genes in the rpoN mutant. The sigma(54)-controlled mannose PTS was shown also to transport glucose in L. plantarum wild-type cells, and its presence causes a lag phase when cultures are transferred from glucose- to galactose-containing media. The mannose PTS appeared to drain phosphoenolpyruvate (PEP) pools in resting cells, since no PEP could be detected in resting wild-type cells, while mannose PTS mutant derivatives contained 1-3 muM PEP (mg protein)(-1 ). Our data provide new insight into the role of sigma( 54) in L. plantarum and possibly other Gram-positive bacteria in the control of expression of an important glucose transporter that contributes to glucose-mediated catabolite control via modulation of the PEP pool.
Stevens
Marc J A
MJA
NIZO food research, PO Box 20, 6710 BA Ede, The Netherlands.|TI Food and Nutrition, PO Box 557, 6700 AN Wageningen, The Netherlands.|Laboratory of Microbiology, Wageningen University and Research Centre, Dreijenplein 10, 6703 HB Wageningen, The Netherlands.
Molenaar
Douwe
D
NIZO food research, PO Box 20, 6710 BA Ede, The Netherlands.|TI Food and Nutrition, PO Box 557, 6700 AN Wageningen, The Netherlands.
de Jong
Anne
A
Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), Rijksuniversiteit Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands.
De Vos
Willem M
WM
Laboratory of Microbiology, Wageningen University and Research Centre, Dreijenplein 10, 6703 HB Wageningen, The Netherlands.|TI Food and Nutrition, PO Box 557, 6700 AN Wageningen, The Netherlands.
Kleerebezem
Michiel
M
Laboratory of Microbiology, Wageningen University and Research Centre, Dreijenplein 10, 6703 HB Wageningen, The Netherlands.|NIZO food research, PO Box 20, 6710 BA Ede, The Netherlands.|TI Food and Nutrition, PO Box 557, 6700 AN Wageningen, The Netherlands.
D050260:Carbohydrate Metabolism; D003470:Culture Media; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D048191:Lactiplantibacillus plantarum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D008358:Mannose / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D009876:Operon; D010728:Phosphoenolpyruvate / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051877:RNA Polymerase Sigma 54 / Q000235:genetics / Q000378:metabolism*; D012329:RNA, Bacterial / Q000235:genetics; D018085:Regulon; D017422:Sequence Analysis, DNA
D003470:Culture Media; D012329:RNA, Bacterial; D010728:Phosphoenolpyruvate; D010770:Phosphotransferases; D051877:RNA Polymerase Sigma 54; D008358:Mannose