The Essential Roles of Cytidine Diphosphate-diacylglycerol Synthase in Bloodstream Form Trypanosoma Brucei

Overview

Journal Mol Microbiol

Specialties Microbiology
Molecular Biology

Date 2014 Feb 19

PMID 24533860

Citations 13

Authors

Alison C Lilley

Louise Major

Simon Young

Michael J R Stark

Terry K Smith

Affiliations

Soon will be listed here.

Abstract

Lipid metabolism in Trypanosoma brucei, the causative agent of African sleeping sickness, differs from its human host in several fundamental ways. This has lead to the validation of a plethora of novel drug targets, giving hope of novel chemical intervention against this neglected disease. Cytidine diphosphate diacylglycerol (CDP-DAG) is a central lipid intermediate for several pathways in both prokaryotes and eukaryotes, being produced by CDP-DAG synthase (CDS). However, nothing is known about the single T. brucei CDS gene (Tb927.7.220/EC 2.7.7.41) or its activity. In this study we show TbCDS is functional by complementation of a non-viable yeast CDS null strain and that it is essential in the bloodstream form of the parasite via a conditional knockout. The TbCDS conditional knockout showed morphological changes including a cell-cycle arrest due in part to kinetoplast segregation defects. Biochemical phenotyping of TbCDS conditional knockout showed drastically altered lipid metabolism where reducing levels of phosphatidylinositol detrimentally impacted on glycoylphosphatidylinositol biosynthesis. These studies also suggest that phosphatidylglycerol synthesized via the phosphatidylglycerol-phosphate synthase is not synthesized from CDP-DAG, as was previously thought. TbCDS was shown to localized the ER and Golgi, probably to provide CDP-DAG for the phosphatidylinositol synthases.

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References

Hammarton T, Lillico S, Welburn S, Mottram J . Trypanosoma brucei MOB1 is required for accurate and efficient cytokinesis but not for exit from mitosis. Mol Microbiol. 2005; 56(1):104-16. PMC: 2244713. DOI: 10.1111/j.1365-2958.2005.04542.x. View

Nickels Jr J, Buxeda R, Carman G . Regulation of phosphatidylinositol 4-kinase from the yeast Saccharomyces cerevisiae by CDP-diacylglycerol. J Biol Chem. 1994; 269(15):11018-24. View

Kuchler K, Daum G, Paltauf F . Subcellular and submitochondrial localization of phospholipid-synthesizing enzymes in Saccharomyces cerevisiae. J Bacteriol. 1986; 165(3):901-10. PMC: 214514. DOI: 10.1128/jb.165.3.901-910.1986. View

Saito S, Goto K, TONOSAKI A, Kondo H . Gene cloning and characterization of CDP-diacylglycerol synthase from rat brain. J Biol Chem. 1997; 272(14):9503-9. DOI: 10.1074/jbc.272.14.9503. View

Smith T, Vasileva N, Gluenz E, Terry S, Portman N, Kramer S . Blocking variant surface glycoprotein synthesis in Trypanosoma brucei triggers a general arrest in translation initiation. PLoS One. 2009; 4(10):e7532. PMC: 2762041. DOI: 10.1371/journal.pone.0007532. View

Roper J, Guther M, Milne K, Ferguson M . Galactose metabolism is essential for the African sleeping sickness parasite Trypanosoma brucei. Proc Natl Acad Sci U S A. 2002; 99(9):5884-9. PMC: 122871. DOI: 10.1073/pnas.092669999. View

Martin K, Smith T . Phosphatidylinositol synthesis is essential in bloodstream form Trypanosoma brucei. Biochem J. 2006; 396(2):287-95. PMC: 1462709. DOI: 10.1042/BJ20051825. View

Nigou J, Besra G . Cytidine diphosphate-diacylglycerol synthesis in Mycobacterium smegmatis. Biochem J. 2002; 367(Pt 1):157-62. PMC: 1222876. DOI: 10.1042/BJ20020370. View

Raetz C, Kennedy E . Function of cytidine diphosphate-diglyceride and deoxycytidine diphosphate-diglyceride in the biogenesis of membrane lipids in Escherichia coli. J Biol Chem. 1973; 248(3):1098-105. View

10.

Shen H, Dowhan W . Reduction of CDP-diacylglycerol synthase activity results in the excretion of inositol by Saccharomyces cerevisiae. J Biol Chem. 1996; 271(46):29043-8. DOI: 10.1074/jbc.271.46.29043. View

11.

Carman G, Kelley M . CDPdiacylglycerol synthase from yeast. Methods Enzymol. 1992; 209:242-7. DOI: 10.1016/0076-6879(92)09030-7. View

12.

Koonin E . A duplicated catalytic motif in a new superfamily of phosphohydrolases and phospholipid synthases that includes poxvirus envelope proteins. Trends Biochem Sci. 1996; 21(7):242-3. View

13.

Ponting C, Kerr I . A novel family of phospholipase D homologues that includes phospholipid synthases and putative endonucleases: identification of duplicated repeats and potential active site residues. Protein Sci. 1996; 5(5):914-22. PMC: 2143407. DOI: 10.1002/pro.5560050513. View

14.

Halford S, Dulai K, Daw S, Fitzgibbon J, Hunt D . Isolation and chromosomal localization of two human CDP-diacylglycerol synthase (CDS) genes. Genomics. 1998; 54(1):140-4. DOI: 10.1006/geno.1998.5547. View

15.

Ganong B, Leonard J, Raetz C . Phosphatidic acid accumulation in the membranes of Escherichia coli mutants defective in CDP-diglyceride synthetase. J Biol Chem. 1980; 255(4):1623-9. View

16.

Felsenstein J . Mathematics vs. Evolution: Mathematical Evolutionary Theory. Science. 1989; 246(4932):941-2. DOI: 10.1126/science.246.4932.941. View

17.

Lykidis A, Jackson P, Rock C, Jackowski S . The role of CDP-diacylglycerol synthetase and phosphatidylinositol synthase activity levels in the regulation of cellular phosphatidylinositol content. J Biol Chem. 1998; 272(52):33402-9. DOI: 10.1074/jbc.272.52.33402. View

18.

Rodgers M, Albanesi J, Phillips M . Phosphatidylinositol 4-kinase III-beta is required for Golgi maintenance and cytokinesis in Trypanosoma brucei. Eukaryot Cell. 2007; 6(7):1108-18. PMC: 1951100. DOI: 10.1128/EC.00107-07. View

19.

Chang S, Heacock P, Clancey C, Dowhan W . The PEL1 gene (renamed PGS1) encodes the phosphatidylglycero-phosphate synthase of Saccharomyces cerevisiae. J Biol Chem. 1998; 273(16):9829-36. DOI: 10.1074/jbc.273.16.9829. View

20.

Johnson J, EDWARDS A, Newton A . A putative phosphatidylserine binding motif is not involved in the lipid regulation of protein kinase C. J Biol Chem. 1998; 272(49):30787-92. DOI: 10.1074/jbc.272.49.30787. View