title: Legionella dumoffii utilizes exogenous choline for phosphatidylcholine synthesis.
abstract: Phosphatidycholine (PC) is the major membrane-forming phospholipid in eukaryotes but it has been found in only a limited number of prokaryotes. Bacteria synthesize PC via the phospholipid N-methylation pathway (Pmt) or via the phosphatidylcholine synthase pathway (Pcs) or both. Here, we demonstrated that Legionella dumoffii has the ability to utilize exogenous choline for phosphatidylcholine (PC) synthesis when bacteria grow in the presence of choline. The Pcs seems to be a primary pathway for synthesis of this phospholipid in L. dumoffii. Structurally different PC species were distributed in the outer and inner membranes. As shown by the LC/ESI-MS analyses, PC15:0/15:0, PC16:0/15:0, and PC17:0/17:1 were identified in the outer membrane and PC14:0/16:0, PC16:0/17:1, and PC20:0/15:0 in the inner membrane. L. dumoffii pcsA gene encoding phosphatidylcholine synthase revealed the highest sequence identity to pcsA of L. bozemanae (82%) and L. longbeachae (81%) and lower identity to pcsA of L. drancourtii (78%) and L. pneumophila (71%). The level of TNF-α in THP1-differentiated cells induced by live and temperature-killed L. dumoffii cultured on a medium supplemented with choline was assessed. Live L. dumoffii bacteria cultured on the choline-supplemented medium induced TNF-α three-fold less efficiently than cells grown on the non-supplemented medium. There is an evident effect of PC modification, which impairs the macrophage inflammatory response.
authors.affiliation: Department of Genetics and Microbiology, Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19 St., 20-033 Lublin, Poland. marta.szysz@poczta.umcs.lublin.pl.
authors.forename: Marta
authors.initials: M
authors.lastname: Palusinska-Szysz
authors.affiliation: Department of Virology and Immunology, Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19 St., 20-033 Lublin, Poland. aszusterciesielska@gmail.com.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Szuster-Ciesielska
authors.affiliation: Mass Spectrometry Group, Institute of Organic Chemistry Polish Academy of Sciences, Kasprzaka 44/52 St., 01-224 Warsaw, Poland. magdalena.kania@icho.edu.pl.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kania
authors.affiliation: Department of Genetics and Microbiology, Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19 St., 20-033 Lublin, Poland. mon.jan@poczta.umcs.lublin.pl.
authors.forename: Monika
authors.initials: M
authors.lastname: Janczarek
authors.affiliation: Department of Genetics and Microbiology, Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19 St., 20-033 Lublin, Poland. elawisniewska87@gmail.com.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Chmiel
authors.affiliation: Mass Spectrometry Group, Institute of Organic Chemistry Polish Academy of Sciences, Kasprzaka 44/52 St., 01-224 Warsaw, Poland. witold.danikiewicz@icho.edu.pl.
authors.forename: Witold
authors.initials: W
authors.lastname: Danikiewicz
mesh_terms: D053898:Biosynthetic Pathways; D002460:Cell Line; D002794:Choline / Q000378:metabolism*; D005798:Genes, Bacterial; D006801:Humans; D007875:Legionella / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D008264:Macrophages / Q000382:microbiology; D013058:Mass Spectrometry; D010713:Phosphatidylcholines / Q000032:analysis / Q000378:metabolism; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics / Q000378:metabolism
chemical_list: D010713:Phosphatidylcholines; D017855:Transferases (Other Substituted Phosphate Groups); C483298:CDP-diacylglycerol choline O-phosphatidyltransferase; D002794:Choline