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Role of CLASP2 in microtubule stabilization and the regulation of persistent motility

机译：CLASP2在微管稳定和持续运动调节中的作用

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In motile fibroblasts, stable microtubules (MTs) are oriented toward the leading edge of cells (1]. How these polarized MT arrays are established and maintained, and the cellular processes they control, have been the subject of many investigations. Several MT "plus-end-tracking proteins," or + TIPs [2], have been proposed to regulate selective MT stabilization, including the CLASPs [3], a complex of CLIP-170, IQGAP1, activated Cdc42 or Rac1 [4], a complex of APC, EB1, and mDia1 [5], and the actin-MT crosslinking factor ACF7 [6]. By using mouse embryonic fibroblasts (MEFs) in a wound-healing assay, we show here that CLASP2 is required for the formation of a stable, polarized MT array but that CLIP-170 and an APC-EB1 interaction are not essential. Persistent motility is also hampered in CLASP2-deficient MEFs. We find that ACF7 regulates cortical CLASP localization in HeLa cells, indicating it acts upstream of CLASP2. Fluorescence-based approaches show that GFP-CLASP2 is immobilized in a bimodal manner in regions near cell edges. Our results suggest that the regional immobilization of CLASP2 allows MT stabilization and promotes directionally persistent motility in fibroblasts.
机译：在活动性成纤维细胞中，稳定的微管（MT）朝向细胞的前缘（1）。如何建立和维持这些极化的MT阵列以及如何控制它们的细胞过程，已成为许多研究的主题。 -末端追踪蛋白，或+ TIPs [2]，已被提议来调节选择性MT的稳定，包括CLASPs [3]，CLIP-170，IQGAP1，激活的Cdc42或Rac1的复合物[4]，CTP-170的复合物。 APC，EB1和mDia1 [5]，以及肌动蛋白-MT交联因子ACF7 [6]。通过在伤口愈合试验中使用小鼠胚胎成纤维细胞（MEF），我们在这里显示CLASP2是形成稳定细胞所必需的，极化的MT阵列，但CLIP-170和APC-EB1的相互作用不是必需的，在缺乏CLASP2的MEF中持久性运动也受到阻碍，我们发现ACF7调节HeLa细胞中皮质CLASP的定位，表明它在CLASP2的上游起作用。基于方法的研究表明，GFP-CLASP2被固定在在细胞边缘附近的区域中双峰方式。我们的结果表明，CLASP2的区域固定可以使MT稳定并促进成纤维细胞的定向持久运动。

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