A role for the dynamic acylation of a cluster of cysteine residues in regulating the activity of the glycosylphosphatidylinositol-specific phospholipase C of Trypanosoma brucei.
The glycosylphosphatidylinositol-specific phospholipase C or VSG lipase is the enzyme responsible for the cleavage of the glycosylphosphatidylinositol anchor of the variant surface glycoprotein (VSG) and concomitant release of the surface coat in Trypanosoma brucei during osmotic shock or extracellular acidic stress. In Xenopus laevis oocytes the VSG lipase was expressed as a nonacylated and a thioacylated form. This thioacylation occurred within a cluster of three cysteine residues but was not essential for catalytic activity per se. These two forms were also detected in trypanosomes and appeared to be present at roughly equivalent amounts. A reversible shift to the acylated form occurred when cells were triggered to release the VSG by either nonlytic acid stress or osmotic lysis. A wild type VSG lipase or a gene mutated in the three codons for the acylated cysteines were reinserted in the genome of a trypanosome null mutant for this gene. A comparative analysis of these revertant trypanosomes indicated that thioacylation might be involved in regulating enzyme access to the VSG substrate.
Paturiaux-Hanocq
F
F
Department of Molecular Biology, Université Libre de Bruxelles, 12 rue des Profs Jeener et Brachet, B-6041, Gosselies, Belgium.
Hanocq-Quertier
J
J
de Almeida
M L
ML
Nolan
D P
DP
Pays
A
A
Vanhamme
L
L
Van den Abbeele
J
J
Wasunna
C L
CL
Carrington
M
M
Pays
E
E
D000215:Acylation; D000818:Animals; D030801:Animals, Genetically Modified; D003545:Cysteine / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D054800:Glycosylphosphatidylinositol Diacylglycerol-Lyase; D016297:Mutagenesis, Site-Directed; D019814:Myristic Acid / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism; D010957:Plasmids; D012333:RNA, Messenger / Q000378:metabolism; D013329:Structure-Activity Relationship; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014162:Transfection; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000235:genetics; D010738:Type C Phospholipases / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis
D012333:RNA, Messenger; D013438:Sulfhydryl Compounds; D019814:Myristic Acid; D010738:Type C Phospholipases; D054800:Glycosylphosphatidylinositol Diacylglycerol-Lyase; D003545:Cysteine