title: Leishmania donovani-induced ceramide as the key mediator of Akt dephosphorylation in murine macrophages: role of protein kinase Czeta and phosphatase.
abstract: Leishmania donovani is an intracellular protozoan parasite that impairs the host macrophage immune response to render it suitable for its survival and establishment. L. donovani-induced immunosuppression and alteration of host cell signaling is mediated by ceramide, a pleiotropic second messenger playing an important role in regulation of several kinases, including mitogen-activated protein kinase and phosphatases. We observed that the endogenous ceramide generated during leishmanial infection led to the dephosphorylation of protein kinase B (PKB) (Akt) in infected cells. The study of ceramide-mediated Akt phosphorylation revealed that Akt was dephosphorylated at both Thr308 and Ser473 sites in infected cells. Further investigation demonstrated that ceramide was also responsible for the induction of PKCzeta, an atypical Ca-independent stress kinase, as well as the ceramide-activated protein phosphatases (e.g., protein phosphatase 2A [PP2A]). We found that Akt dephosphorylation was mediated by ceramide-induced PKCzeta-Akt association and PP2A activation. In addition, treatment of L. donovani-infected macrophages with PKCzeta-specific inhibitor peptide could restore the translocation of phosphorylated Akt to the cell membrane. This study also revealed that ceramide is involved in the inhibition of proinflammatory cytokine tumor necrosis factor alpha release by infected macrophages. These observations strongly suggest the importance of ceramide in the alteration of normal cellular functions, impairment of the kinase/phosphatase balance, and thereby establishment of leishmaniasis in the hostile macrophage environment.
authors.affiliation: Dept. of Microbiology, Bose Institute, P-1/12, CIT Scheme VII M, Kolkata, India.
authors.forename: Ranadhir
authors.initials: R
authors.lastname: Dey
authors.forename: Nivedita
authors.initials: N
authors.lastname: Majumder
authors.forename: Surajit
authors.initials: S
authors.lastname: Bhattacharjee
authors.forename: Suchandra Bhattacharyya
authors.initials: SB
authors.lastname: Majumdar
authors.forename: Rajdeep
authors.initials: R
authors.lastname: Banerjee
authors.forename: Sandipan
authors.initials: S
authors.lastname: Ganguly
authors.forename: Pradeep
authors.initials: P
authors.lastname: Das
authors.forename: Subrata
authors.initials: S
authors.lastname: Majumdar
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002518:Ceramides / Q000378:metabolism / Q000494:pharmacology*; D004789:Enzyme Activation; D005260:Female; D007893:Leishmania donovani / Q000472:pathogenicity*; D017737:Macrophages, Peritoneal / Q000201:enzymology / Q000469:parasitology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D054648:Protein Phosphatase 2; D051057:Proto-Oncogene Proteins c-akt / Q000737:chemistry / Q000378:metabolism*; D000099221:Protein Kinase C zeta
chemical_list: D002518:Ceramides; D010749:Phosphoprotein Phosphatases; D011493:Protein Kinase C; D054648:Protein Phosphatase 2; D051057:Proto-Oncogene Proteins c-akt; D000099221:Protein Kinase C zeta