Center for Infectious Disease Research, Seattle, WA 98109.
Department of Global Health, University of Washington, Seattle, WA 98195.
Mol Biol Cell. 2018 May 1;29(9):1137-1152. doi: 10.1091/mbc.E17-08-0515. Epub 2018 Mar 22.
Many cellular processes change during the Trypanosoma brucei life cycle as this parasite alternates between the mammalian host and tsetse fly vector. We show that the inositol phosphate pathway helps regulate these developmental changes. Knockdown of inositol polyphosphate multikinase (IPMK), which phosphorylates Ins(1,4,5)P3 and Ins(1,3,4,5)P4, resulted in changes in bloodstream forms that are characteristic of insect stage procyclic forms. These changes include expression of the procyclic surface coat, up-regulation of RNA-binding proteins that we show to regulate stage-specific transcripts, and activation of oxidative phosphorylation with increased ATP production in bloodstream forms. These changes were accompanied by development of procyclic morphology, which also occurred by the expression of a catalytically inactive IPMK, implying that regulation of these processes entails IPMK activity. Proteins involved in signaling, protein synthesis and turnover, and metabolism were affinity-enriched with the IPMK substrate or product. Developmental changes associated with IPMK knockdown or catalytic inactivation reflected processes that are enriched with inositol phosphates, and chemical and genetic perturbation of these processes affected T. brucei development. Hence, IPMK helps regulate T. brucei development, perhaps by affecting inositol phosphate interactions with proteins of the regulatory network that controls energy metabolism and development.
许多细胞过程在布氏锥虫生命周期中发生变化,因为这种寄生虫在哺乳动物宿主和采采蝇媒介之间交替。我们表明,肌醇磷酸盐途径有助于调节这些发育变化。肌醇多磷酸激酶(IPMK)的敲低,其磷酸化 Ins(1,4,5)P3 和 Ins(1,3,4,5)P4,导致血液形式的变化,这些变化是昆虫阶段前鞭毛体形式的特征。这些变化包括前鞭毛体表面被覆蛋白的表达,上调我们表明调节阶段特异性转录物的 RNA 结合蛋白,以及氧化磷酸化的激活,导致血液形式中 ATP 产生增加。这些变化伴随着前鞭毛体形态的发育,这也通过表达无催化活性的 IPMK 发生,这意味着这些过程的调节需要 IPMK 活性。参与信号转导、蛋白质合成和周转以及代谢的蛋白质与 IPMK 底物或产物进行了亲和富集。与 IPMK 敲低或催化失活相关的发育变化反映了富含肌醇磷酸盐的过程,这些过程的化学和遗传扰动影响了 T. brucei 的发育。因此,IPMK 有助于调节 T. brucei 的发育,也许是通过影响肌醇磷酸盐与调控能量代谢和发育的调控网络中的蛋白质相互作用。