title: Elucidation and chemical modulation of sulfolipid-1 biosynthesis in Mycobacterium tuberculosis.
abstract: Mycobacterium tuberculosis possesses unique cell-surface lipids that have been implicated in virulence. One of the most abundant is sulfolipid-1 (SL-1), a tetraacyl-sulfotrehalose glycolipid. Although the early steps in SL-1 biosynthesis are known, the machinery underlying the final acylation reactions is not understood. We provide genetic and biochemical evidence for the activities of two proteins, Chp1 and Sap (corresponding to gene loci rv3822 and rv3821), that complete this pathway. The membrane-associated acyltransferase Chp1 accepts a synthetic diacyl sulfolipid and transfers an acyl group regioselectively from one donor substrate molecule to a second acceptor molecule in two successive reactions to yield a tetraacylated product. Chp1 is fully active in vitro, but in M. tuberculosis, its function is potentiated by the previously identified sulfolipid transporter MmpL8. We also show that the integral membrane protein Sap and MmpL8 are both essential for sulfolipid transport. Finally, the lipase inhibitor tetrahydrolipstatin disrupts Chp1 activity in M. tuberculosis, suggesting an avenue for perturbing SL-1 biosynthesis in vivo. These data complete the SL-1 biosynthetic pathway and corroborate a model in which lipid biosynthesis and transmembrane transport are coupled at the membrane-cytosol interface through the activity of multiple proteins, possibly as a macromolecular complex.
authors.affiliation: Department of Chemistry, University of California, Berkeley, California 94720-1460, USA.
authors.forename: Jessica C
authors.initials: JC
authors.lastname: Seeliger
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Holsclaw
authors.forename: Michael W
authors.initials: MW
authors.lastname: Schelle
authors.forename: Zsofia
authors.initials: Z
authors.lastname: Botyanszki
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Gilmore
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Tully
authors.forename: Michael
authors.initials: M
authors.lastname: Niederweis
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
authors.forename: Julie A
authors.initials: JA
authors.lastname: Leary
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Bertozzi
mesh_terms: D000215:Acylation / Q000502:physiology; D000217:Acyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D001693:Biological Transport, Active / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006017:Glycolipids / Q000096:biosynthesis* / Q000235:genetics; D007783:Lactones / Q000494:pharmacology; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism*; D000077403:Orlistat; D037521:Virulence Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D006017:Glycolipids; D007783:Lactones; D037521:Virulence Factors; C009355:sulfolipid I; D000077403:Orlistat; D000217:Acyltransferases