Cellular Requirements for the Small Molecule Forchlorfenuron to Stabilize the Septin Cytoskeleton
Cellular Requirements for the Small Molecule Forchlorfenuron to Stabilize the Septin Cytoskeleton
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DOI：
10.1002/cm.20452
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发表时间：
2010-06-01
期刊：
影响因子：
2.9
通讯作者：
Gladfelter, Amy S.
中科院分区：
文献类型：
--
作者：
DeMay, Bradley S.;Meseroll, Rebecca A.;Gladfelter, Amy S.
septins是一种能形成葡萄糖的GTP结合蛋白，从酵母到人类都是保守的。隔膜蛋白组装成更高级的结构，如环，棒和纱布，具有多种功能，包括作为膜扩散屏障和细胞信号传导的支架。Septin长丝聚合的基础和Septin聚合物动力学的规则目前还不清楚。药理学试剂是研究肌动蛋白和微管细胞骨架的这些特性的基本工具，然而只有有限的报道特异性针对隔蛋白细胞骨架的药物。氯吡脲（FCF）是一种用于农业的合成植物细胞分裂素，已被证明可以改变酵母和哺乳动物组织培养细胞中的Septin组织。在这里，我们评估细胞的要求和性质的septin为基础的结构诱导FCF。用FCF处理丝状真菌棉阿舒囊菌导致广泛的隔蛋白纤维在整个菌丝中组装，这在去除药物后迅速逆转。这些纤维在组装后不交换或添加隔蛋白亚基，表明FCF抑制正常的隔蛋白动力学并稳定聚合物。虽然FCF诱导的隔蛋白纤维不共定位于肌动蛋白或微管，但极化的F-肌动蛋白细胞骨架可能需要药物诱导的隔蛋白纤维的组装。因此，FCF是隔蛋白聚合的有效诱导剂，并且充当延长的隔蛋白聚合物的可逆稳定剂。这种药物将成为研究septin聚合和功能机制的有力工具，特别是在分子分析因存在多种亚型和有限遗传学而复杂的细胞类型中。(C)2010 Wiley-Liss，Inc
The septins are filament-forming, GTP-binding proteins that are conserved from yeast to humans. Septins assemble into higher-order structures such as rings, bars, and gauzes with diverse functions including serving as membrane diffusion barriers and scaffolds for cell signaling. The basis for septin filament polymerization and the rules governing septin polymer dynamics are presently not well understood. Pharmacological agents are essential tools in studying such properties of the actin and microtubule cytoskeletons however there are only limited reports of a drug specific to the septin cytoskeleton. Forchlorfenuron (FCF) is a synthetic plant cytokinin used in agriculture which has been shown to alter septin organization in yeast and mammalian tissue culture cells. Here we assess cellular requirements and properties of septin-based structures induced by FCF. Treatment of the filamentous fungus Ashbya gossypii with FCF leads to assembly of extensive septin fibers throughout hyphae which is rapidly reversed upon removal of the drug. These fibers do not exchange or add septin subunits after assembly, indicating that FCF suppresses normal septin dynamics and stabilizes the polymers. While FCF-induced septin fibers do not co-localize to actin or microtubules, a polarized F-actin cytoskeleton is likely required for the assembly of drug-induced septin fibers. Thus, FCF is a potent inducer of septin polymerization and acts as a reversible stabilizer of extended septin polymers. This drug will be a powerful tool for studying mechanisms of septin polymerization and function, particularly in cell types where molecular analyses are complicated by the presence of multiple isoforms and limited genetics. (C) 2010 Wiley-Liss, Inc