title: 1,2-Diacylglycerol choline phosphotransferase catalyzes the final step in the unique Treponema denticola phosphatidylcholine biosynthesis pathway.
abstract: Treponema denticola synthesizes phosphatidylcholine through a licCA-dependent CDP-choline pathway identified only in the genus Treponema. However, the mechanism of conversion of CDP-choline to phosphatidylcholine remained unclear. We report here characterization of TDE0021 (herein designated cpt) encoding a 1,2-diacylglycerol choline phosphotransferase homologous to choline phosphotransferases that catalyze the final step of the highly conserved Kennedy pathway for phosphatidylcholine synthesis in eukaryotes. T. denticola Cpt catalyzed in vitro phosphatidylcholine formation from CDP-choline and diacylglycerol, and full activity required divalent manganese. Allelic replacement mutagenesis of cpt in T. denticola resulted in abrogation of phosphatidylcholine synthesis. T. denticola Cpt complemented a Saccharomyces cerevisiae CPT1 mutant, and expression of the entire T. denticola LicCA-Cpt pathway in E. coli resulted in phosphatidylcholine biosynthesis. Our findings show that T. denticola possesses a unique phosphatidylcholine synthesis pathway combining conserved prokaryotic choline kinase and CTP:phosphocholine cytidylyltransferase activities with a 1,2-diacylglycerol choline phosphotransferase that is common in eukaryotes. Other than in a subset of mammalian host-associated Treponema that includes T. pallidum, this pathway is found in neither bacteria nor Archaea. Molecular dating analysis of the Cpt gene family suggests that a horizontal gene transfer event introduced this gene into an ancestral Treponema well after its divergence from other spirochetes.
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av. Universidad s/n, Apdo. Postal 565-A, Cuernavaca, Morelos, CP62210, Mexico.
authors.forename: Miguel Ángel
authors.initials: MÁ
authors.lastname: Vences-Guzmán
authors.affiliation: Department of Biologic and Materials Sciences, University of Michigan School of Dentistry, Ann Arbor, Michigan, 48109, USA.
authors.forename: M
authors.initials: M
authors.lastname: Paula Goetting-Minesky
authors.affiliation: Department of Biochemistry, Duke University Medical Center, Durham, North Carolina, 27710, USA.
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Guan
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av. Universidad s/n, Apdo. Postal 565-A, Cuernavaca, Morelos, CP62210, Mexico.
authors.forename: Santiago
authors.initials: S
authors.lastname: Castillo-Ramirez
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av. Universidad s/n, Apdo. Postal 565-A, Cuernavaca, Morelos, CP62210, Mexico.
authors.forename: Luz América
authors.initials: LA
authors.lastname: Córdoba-Castro
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av. Universidad s/n, Apdo. Postal 565-A, Cuernavaca, Morelos, CP62210, Mexico.
authors.forename: Isabel M
authors.initials: IM
authors.lastname: López-Lara
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av. Universidad s/n, Apdo. Postal 565-A, Cuernavaca, Morelos, CP62210, Mexico.
authors.forename: Otto
authors.initials: O
authors.lastname: Geiger
authors.affiliation: Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av. Universidad s/n, Apdo. Postal 565-A, Cuernavaca, Morelos, CP62210, Mexico.
authors.forename: Christian
authors.initials: C
authors.lastname: Sohlenkamp
authors.affiliation: Department of Biologic and Materials Sciences, University of Michigan School of Dentistry, Ann Arbor, Michigan, 48109, USA.
authors.forename: J
authors.initials: J
authors.lastname: Christopher Fenno
mesh_terms: D000483:Alleles; D053898:Biosynthetic Pathways / Q000235:genetics / Q000502:physiology; D002384:Catalysis; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism*; D007700:Kinetics; D008345:Manganese / Q000378:metabolism; D016296:Mutagenesis; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D016415:Sequence Alignment; D048373:Treponema denticola / Q000235:genetics / Q000378:metabolism*
chemical_list: D010713:Phosphatidylcholines; D008345:Manganese; D002798:Diacylglycerol Cholinephosphotransferase