Multiple polarity kinases inhibit phase separation of F-BAR protein Cdc15 and antagonize cytokinetic ring assembly in fission yeast.
Multiple polarity kinases inhibit phase separation of F-BAR protein Cdc15 and antagonize cytokinetic ring assembly in fission yeast.
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DOI：
10.7554/elife.83062
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发表时间：
2023-02-07
期刊：
影响因子：
7.7
通讯作者：
Gould KL
中科院分区：
文献类型：
--
作者：
Bhattacharjee R;Hall AR;Mangione MC;Igarashi MG;Roberts-Galbraith RH;Chen JS;Vavylonis D;Gould KL
F-BAR蛋白Cdc 15是粟酒裂殖酵母胞质分裂所必需的蛋白，在将细胞动力学环（CR）附着到质膜（PM）上起关键作用。Cdc 15通过其F-BAR结构域与膜结合和寡聚化的能力受到其固有无序区（IDR）磷酸化的抑制。多种细胞极性激酶调节Cdc 15 IDR磷酸化，其中Cdc 15上的DYRK激酶Pom 1磷酸化位点已在体内显示出阻止细胞尖端的CR形成。在这里，我们比较了Pom 1控制Cdc 15磷酸化和皮质定位的能力，其他Cdc 15激酶：Kin 1，Pck 1和Shk 1。我们确定了不同的，但重叠的队列Cdc 15磷酸化位点的每一个激酶的目标，和网站的数量与每一个激酶的能力，影响Cdc 15 PM定位。粗粒度模拟预测，累积的IDR磷酸化移动的IDR的二聚体分开，并向F-BAR的提示。此外，模拟表明磷酸化的总体负电荷掩盖了F-BAR寡聚化和膜相互作用所需的带正电的氨基酸。最后，模拟表明，去磷酸化Cdc 15经历IDR相互作用驱动的相分离。事实上，去磷酸化但未磷酸化的Cdc 15经历液-液相分离以在体外形成招募Cdc 15结合配偶体的液滴。在细胞中，Cdc 15磷酸突变体也形成了PM结合的凝聚物，招募其他CR组件。总之，我们建议，由各种激酶的Cdc 15磷酸化的阈值防止Cdc 15的PM上的冷凝和拮抗CR组装。
The F-BAR protein Cdc15 is essential for cytokinesis in Schizosaccharomyces pombe and plays a key role in attaching the cytokinetic ring (CR) to the plasma membrane (PM). Cdc15’s abilities to bind to the membrane and oligomerize via its F-BAR domain are inhibited by phosphorylation of its intrinsically disordered region (IDR). Multiple cell polarity kinases regulate Cdc15 IDR phosphostate, and of these the DYRK kinase Pom1 phosphorylation sites on Cdc15 have been shown in vivo to prevent CR formation at cell tips. Here, we compared the ability of Pom1 to control Cdc15 phosphostate and cortical localization to that of other Cdc15 kinases: Kin1, Pck1, and Shk1. We identified distinct but overlapping cohorts of Cdc15 phosphorylation sites targeted by each kinase, and the number of sites correlated with each kinases’ abilities to influence Cdc15 PM localization. Coarse-grained simulations predicted that cumulative IDR phosphorylation moves the IDRs of a dimer apart and toward the F-BAR tips. Further, simulations indicated that the overall negative charge of phosphorylation masks positively charged amino acids necessary for F-BAR oligomerization and membrane interaction. Finally, simulations suggested that dephosphorylated Cdc15 undergoes phase separation driven by IDR interactions. Indeed, dephosphorylated but not phosphorylated Cdc15 undergoes liquid–liquid phase separation to form droplets in vitro that recruit Cdc15 binding partners. In cells, Cdc15 phosphomutants also formed PM-bound condensates that recruit other CR components. Together, we propose that a threshold of Cdc15 phosphorylation by assorted kinases prevents Cdc15 condensation on the PM and antagonizes CR assembly.
影响因子：
5.6
作者：
Smith H;Pinkerton N;Heisler DB;Kudryashova E;Hall AR;Karch KR;Norris A;Wysocki V;Sotomayor M;Reisler E;Vavylonis D;Kudryashov DS
通讯作者：
Kudryashov DS