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The essential roles of cytidine diphosphate‐diacylglycerol synthase in bloodstream form T rypanosoma brucei
Author(s) -
Lilley Alison C.,
Major Louise,
Young Simon,
Stark Michael J. R.,
Smith Terry K.
Publication year - 2014
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/mmi.12553
Subject(s) - trypanosoma brucei , biology , diacylglycerol kinase , phosphatidylinositol , phosphatidylglycerol , complementation , biochemistry , atp synthase , phosphatidic acid , cytidine , saccharomyces cerevisiae , biosynthesis , microbiology and biotechnology , enzyme , yeast , phospholipid , gene , signal transduction , phenotype , protein kinase c , membrane , phosphatidylcholine
Summary Lipid metabolism in T rypanosoma 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 ( Tb 927.7.220/ 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 G olgi, probably to provide CDP‐DAG for the phosphatidylinositol synthases.