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Journal of Bacteriology, June 2001, p. 3391-3398, Vol. 183, No. 11
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.11.3391-3398.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Regulatory Functions of Serine-46-Phosphorylated HPr in Lactococcus lactis

Vicente Monedero,1,dagger Oscar P. Kuipers,2,Dagger Emmanuel Jamet,3 and Josef Deutscher1,*

Laboratoire de Génétique des Microorganismes, INRA-CNRS URA 1925, 78850 Thiverval-Grignon,1 and Génétique Microbienne, INRA, 78352 Jouy en Josas, France,3 and NIZO Food Research, 6710 BA Ede, The Netherlands2

Received 18 December 2000/Accepted 19 March 2001

In most low-G+C gram-positive bacteria, the phosphoryl carrier protein HPr of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) becomes phosphorylated at Ser-46. This ATP-dependent reaction is catalyzed by the bifunctional HPr kinase/P-Ser-HPr phosphatase. We found that serine-phosphorylated HPr (P-Ser-HPr) of Lactococcus lactis participates not only in carbon catabolite repression of an operon encoding a beta -glucoside-specific EII and a 6-P-beta -glucosidase but also in inducer exclusion of the non-PTS carbohydrates maltose and ribose. In a wild-type strain, transport of these non-PTS carbohydrates is strongly inhibited by the presence of glucose, whereas in a ptsH1 mutant, in which Ser-46 of HPr is replaced with an alanine, glucose had lost its inhibitory effect. In vitro experiments carried out with L. lactis vesicles had suggested that P-Ser-HPr is also implicated in inducer expulsion of nonmetabolizable homologues of PTS sugars, such as methyl beta -D-thiogalactoside (TMG) and 2-deoxy-D-glucose (2-DG). In vivo experiments with the ptsH1 mutant established that P-Ser-HPr is not necessary for inducer expulsion. Glucose-activated 2-DG expulsion occurred at similar rates in wild-type and ptsH1 mutant strains, whereas TMG expulsion was slowed in the ptsH1 mutant. It therefore seems that P-Ser-HPr is not essential for inducer expulsion but that in certain cases it can play an indirect role in this regulatory process.

* Corresponding author. Mailing address: Laboratoire de Génétique des Microorganismes, INRA-CNRS URA 1925, 78850 Thiverval-Grignon, France. Phone: 33-1-30815447. Fax: 33-1-30815457. E-mail: jdeu{at}grignon.inra.fr.

dagger Present address: Departamento de Biotecnologia, Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Valencia, Spain.

Dagger Present address: Department of Genetics, University of Groningen, 9751 NN Haren, The Netherlands.

Journal of Bacteriology, June 2001, p. 3391-3398, Vol. 183, No. 11
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.11.3391-3398.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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