Interaction with Enzyme IIB^sup Mpo^ (EIIB^sup Mpo^) and Phosphorylation by Phosphorylated EIIB^sup Mpo^ Exert Antagonistic Effects on the Transcriptional Activator ManR of Listeria monocytogenes

• Published in: Journal of bacteriology 2015-05, Vol.197 (9), p.1559
• Main Authors: Zébré, Arthur Constant, Aké, Francine Moussan, Ventroux, Magali, Koffi-Nevry, Rose, Noirot-Gros, Marie-Françoise, Deutscher, Josef, Milohanic, Eliane
• Format: Article
• Language: English
• Subjects: Bacteria; Bacteriology; Carbohydrates; Effects; Enzymes; Experiments; Phosphorylation; Transcription factors; Yeast; Yeasts
• ISSN: 0021-9193; 1098-5530

Listeriae take up glucose and mannose predominantly through a mannose class phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS...), whose three components are encoded by the manLMN genes. The expression of these genes is controlled by ManR, a LevR-type transcription activator containing two PTS regulation domains (PRDs) and two PTS-like domains (enzyme IIA... [EIIA...]- and EIIB...-like). We demonstrate here that in Listeria monocytogenes, ManR is activated via the phosphorylation of His585 in the EIIA...-like domain by the general PTS components enzyme I and HPr. We also show that ManR is regulated by the PTS... and that EIIB... plays a dual role in ManR regulation. First, yeast two-hybrid experiments revealed that unphosphorylated EIIB... interacts with the two C-terminal domains of ManR (EIIB...-like and PRD2) and that this interaction is required for ManR activity. Second, in the absence of glucose/mannose, phosphorylated EIIB... (P~EIIB...) inhibits ManR activity by phosphorylating His871 in PRD2. The presence of glucose/mannose causes the dephosphorylation of P~EIIB... and P~PRD2 of ManR, which together lead to the induction of the manLMN operon. Complementation of a ...manR mutant with various manR alleles confirmed the antagonistic effects of PTS-catalyzed phosphorylation at the two different histidine residues of ManR. Deletion of manR prevented not only the expression of the manLMN operon but also glucose-mediated repression of virulence gene expression; however, repression by other carbohydrates was unaffected. Interestingly, the expression of manLMN in Listeria innocua was reported to require not only ManR but also the Crp-like transcription activator Lin0142. Unlike Lin0142, the L. monocytogenes homologue, Lmo0095, is not required for manLMN expression; its absence rather stimulates man expression. (ProQuest: ... denotes formulae/symbols omitted.)