Clemmens Clarice S, Morris Meredith T, Lyda Todd A, Acosta-Serrano Alvaro, Morris James C
Department of Genetics and Biochemistry, Clemson University, Clemson, SC 29634, USA.
Exp Parasitol. 2009 Nov;123(3):250-7. doi: 10.1016/j.exppara.2009.07.010. Epub 2009 Aug 6.
The African trypanosome, Trypanosoma brucei, can gauge its environment by sensing nutrient availability. For example, procyclic form (PF) trypanosomes monitor changes in glucose levels to regulate surface molecule expression, which is important for survival in the tsetse fly vector. The molecular connection between glycolysis and surface molecule expression is unknown. Here we partially characterize T. brucei homologs of the beta and gamma subunits of the AMP-activated protein kinase (AMPK), and determine their roles in regulating surface molecule expression. Using flow cytometry and mass spectrometry, we found that TbAMPKbeta or TbAMPKgamma-deficient parasites express both of the major surface molecules, EP- and GPEET-procyclin, with the latter being a form that is expressed when glucose is low such as in the tsetse fly. Last, we have found that the putative scaffold component of the complex, TbAMPKbeta, fractionates with organellar components and colocalizes in part with a glycosomal marker as well as the flagellum of PF parasites.
非洲锥虫,即布氏锥虫,能够通过感知营养物质的可利用性来评估其环境。例如,前循环型(PF)锥虫监测葡萄糖水平的变化以调节表面分子的表达,这对于在采采蝇媒介中生存至关重要。糖酵解与表面分子表达之间的分子联系尚不清楚。在这里,我们部分表征了AMP活化蛋白激酶(AMPK)的β和γ亚基的布氏锥虫同源物,并确定了它们在调节表面分子表达中的作用。使用流式细胞术和质谱分析,我们发现缺乏TbAMPKβ或TbAMPKγ的寄生虫表达两种主要的表面分子,即EP-和GPEET-前环素,后者是在葡萄糖含量低时(如在采采蝇中)表达的一种形式。最后,我们发现该复合物的假定支架成分TbAMPKβ与细胞器成分分离,并部分与糖体标记物以及PF寄生虫的鞭毛共定位。