Leishmania dual-specificity tyrosine-regulated kinase 1 (DYRK1) is required for sustaining Leishmania stationary phase phenotype
Leishmania dual-specificity tyrosine-regulated kinase 1 (DYRK1) is required for sustaining Leishmania stationary phase phenotype
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DOI：
10.1111/mmi.14464
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发表时间：
2020-02-11
影响因子：
3.6
通讯作者：
Smirlis, Despina
中科院分区：
文献类型：
--
作者：
Costa Rocha, Vinicius Pinto;Dacher, Mariko;Smirlis, Despina
虽然倍增和生长停滞状态在利什曼原虫发育中起着关键作用，但这些转变的调节剂在很大程度上是未知的。为了更好地理解这些过程，我们表征了具有双重特异性的蛋白激酶家族的一个成员，LinDYRK 1，它在其他生物中充当停滞调节剂。LinDYRK 1过表达的寄生虫表现出增殖减少和停滞细胞的细胞周期重新进入。缺乏LinDYRK 1的寄生虫在对数生长期和稳定生长期表现出不同的适应性表型。在对数生长期，LinDYRK 1（-/-）寄生虫比对照系增殖得更好，支持这种激酶在停滞中的作用，而在静止生长期，LinDYRK 1（-/-）寄生虫具有重要的缺陷，因为它们聚集，积累空泡和脂质体，并显示出微妙但一致的脂质组成差异。此外，它们表达较少的富含代谢环的转录物，显示对补体裂解的敏感性增加，并且在腹腔巨噬细胞内存活率显著降低。不同的LinDYRK 1（-/-）生长期表型反映了不同的LinDYRK 1定位在对数（主要是在鞭毛口袋区和内体）和后期稳定期（Ephon）。总的来说，这项工作提供了第一个DYRK家族成员在维持前鞭毛体稳定期表型和感染性的作用的证据。
Although the multiplicative and growth-arrested states play key roles in Leishmania development, the regulators of these transitions are largely unknown. In an attempt to gain a better understanding of these processes, we characterised one member of a family of protein kinases with dual specificity, LinDYRK1, which acts as a stasis regulator in other organisms. LinDYRK1 overexpressing parasites displayed a decrease in proliferation and in cell cycle re-entry of arrested cells. Parasites lacking LinDYRK1 displayed distinct fitness phenotypes in logarithmic and stationary growth phases. In logarithmic growth phase, LinDYRK1(-/-) parasites proliferated better than control lines, supporting a role of this kinase in stasis, while in stationary growth phase, LinDYRK1(-/-) parasites had important defects as they rounded up, accumulated vacuoles and lipid bodies and displayed subtle but consistent differences in lipid composition. Moreover, they expressed less metacyclic-enriched transcripts, displayed increased sensitivity to complement lysis and a significant reduction in survival within peritoneal macrophages. The distinct LinDYRK1(-/-) growth phase phenotypes were mirrored by the distinct LinDYRK1 localisations in logarithmic (mainly in flagellar pocket area and endosomes) and late stationary phase (mitochondrion). Overall, this work provides first evidence for the role of a DYRK family member in sustaining promastigote stationary phase phenotype and infectivity.