Plasmodium berghei kinesin-5 associates with the spindle apparatus during cell division and is important for efficient production of infectious sporozoites
Plasmodium berghei kinesin-5 associates with the spindle apparatus during cell division and is important for efficient production of infectious sporozoites
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伯氏疟原虫驱动蛋白-5 在细胞分裂过程中与纺锤体结合，对于有效产生感染性子孢子非常重要
DOI：
10.1101/2020.07.03.186031
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Zeeshan M
中科院分区：
文献类型：
--
作者：
Zeeshan M
在细胞分裂过程中，Kinesin-5马达通过产生力量来建立和维持纺锤体两极，从而在细胞分裂过程中扮演着重要的角色，这是染色体正确分离所必需的。在模式真核生物中，Kinesin-5在结构和功能上基本保守，但它在疟原虫中的作用尚不清楚。疟原虫是一种进化分化的生物，具有有丝分裂和减数分裂的几个非典型特征。我们研究了驱动蛋白-5在伯氏疟原虫整个生命周期的细胞分裂过程中的功能和亚细胞定位。除卵囊产生子孢子外，Kinesin-5的缺失在任何增殖期都没有明显影响，导致蚊子唾液腺中能够感染新的脊椎动物宿主的活动子孢子的数量显著减少。活细胞成像显示，在非典型有丝分裂和减数分裂过程中，Kinesin-5-GFP位于纺锤体和纺锤体两极。固定细胞免疫荧光分析显示，在血液分裂早期，Kinesin-5与α-微管蛋白和Centin-2共定位，并与动粒标记NDC80部分重叠。双色活细胞成像显示，在雄配子发育过程中，Kinesin-5与NDC80密切相关，但与Kinesin-8B无关，Kinesin-8B是形成鞭毛的基底和轴丝的标志。用微管特异性抑制剂处理配子细胞证实了Kinesin-5与核纺锤体有关，而不是与细胞质轴线微管有关。总之，我们的结果表明，Kinesin-5与纺锤体相关，在寄生虫的增殖期表达，对于有效生产感染性子孢子是重要的。
Kinesin-5 motors play essential roles in spindle apparatus assembly during cell division, by generating forces to establish and maintain the spindle bipolarity essential for proper chromosome segregation. Kinesin-5 is largely conserved structurally and functionally in model eukaryotes, but its role is unknown in thePlasmodiumparasite, an evolutionarily divergent organism with several atypical features of both mitotic and meiotic cell division. We have investigated the function and subcellular location of kinesin-5 during cell division throughout thePlasmodium bergheilife cycle. Deletion ofkinesin-5had little visible effect at any proliferative stage except sporozoite production in oocysts, resulting in a significant decrease in the number of motile sporozoites in mosquito salivary glands, which were able to infect a new vertebrate host. Live-cell imaging showed kinesin-5-GFP located on the spindle and at spindle poles during both atypical mitosis and meiosis. Fixed-cell immunofluorescence assays revealed kinesin-5 co-localized with α-tubulin and centrin-2 and a partial overlap with kinetochore marker NDC80 during early blood stage schizogony. Dual-color live-cell imaging showed that kinesin-5 is closely associated with NDC80 during male gametogony, but not with kinesin-8B, a marker of the basal body and axonemes of the forming flagella. Treatment of gametocytes with microtubule-specific inhibitors confirmed kinesin-5 association with nuclear spindles and not cytoplasmic axonemal microtubules. Altogether, our results demonstrate that kinesin-5 is associated with the spindle apparatus, expressed in proliferating parasite stages, and important for efficient production of infectious sporozoites.