Webb H, Carnall N, Vanhamme L, Rolin S, Van Den Abbeele J, Welburn S, Pays E, Carrington M
Department of Biochemistry, Cambridge University, United Kingdom.
J Cell Biol. 1997 Oct 6;139(1):103-14. doi: 10.1083/jcb.139.1.103.
In the mammalian host, the cell surface of Trypanosoma brucei is protected by a variant surface glycoprotein that is anchored in the plasma membrane through covalent attachment of the COOH terminus to a glycosylphosphatidylinositol. The trypanosome also contains a phospholipase C (GPI-PLC) that cleaves this anchor and could thus potentially enable the trypanosome to shed the surface coat of VSG. Indeed, release of the surface VSG can be observed within a few minutes on lysis of trypanosomes in vitro. To investigate whether the ability to cleave the membrane anchor of the VSG is an essential function of the enzyme in vivo, a GPI-PLC null mutant trypanosome has been generated by targeted gene deletion. The mutant trypanosomes are fully viable; they can go through an entire life cycle and maintain a persistent infection in mice. Thus the GPI-PLC is not an essential activity and is not necessary for antigenic variation. However, mice infected with the mutant trypanosomes have a reduced parasitemia and survive longer than those infected with control trypanosomes. This phenotype is partially alleviated when the null mutant is modified to express low levels of GPI-PLC.
在哺乳动物宿主体内,布氏锥虫的细胞表面由一种可变表面糖蛋白保护,该糖蛋白通过其羧基末端与糖基磷脂酰肌醇的共价连接而锚定在质膜上。锥虫还含有一种磷脂酶C(GPI-PLC),它能切割这种锚定物,因此有可能使锥虫脱落VSG表面 coat。事实上,在体外锥虫裂解后几分钟内就能观察到表面VSG的释放。为了研究切割VSG膜锚定物的能力在体内是否是该酶的一项基本功能,通过靶向基因缺失产生了一种GPI-PLC基因敲除突变型锥虫。突变型锥虫完全可以存活;它们能够经历完整的生命周期,并在小鼠体内维持持续性感染。因此,GPI-PLC不是一项基本活性,对抗原变异也不是必需的。然而,感染突变型锥虫小鼠的寄生虫血症低于感染对照锥虫的小鼠,且存活时间更长。当基因敲除突变体经改造后表达低水平的GPI-PLC时,这种表型会得到部分缓解。