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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Characterisation of the Rac/PAK pathway in Amoeba proteus
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Characterisation of the Rac/PAK pathway in Amoeba proteus

机译：变形虫中Rac / PAK途径的表征

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Molecular mechanisms underlying the unique locomotion of the highly motile Amoeba proteus still remain poorly understood. Recently, we have shown that blocking the endogenous amoebal Rac-like protein(s) leads to distinct and irreversible changes in the appearance of these large migrating cells as well as to a significant inhibition of their locomotion. To elucidate the mechanism of the Rac pathway in Amoeba proteus, we tested the effects of blocking the endogenous myosin I heavy chain kinase (MIHCK), one of the Rac effectors in Acanthamoeba castellanii and Dictyostelium discoideum, with anti-MIHCK antibodies in migrating amoebae, as well as the effect of inhibiting Rac and MIHCK on the actin polymerisation process. Antibodies against A. castellanii MIHCK detected an A. proteus protein with a molecular mass (ca. 95 kDa) similar to the A. castellanii kinase. The cellular distribution of MIHCK in A. proteus was very similar to those of Rac-like protein in amoebae and MIHCK in A. castellanii. Amoebae microinjected with anti-MIHCK antibodies moved slower and protruded fewer wide pseudopodia (5-6) than the control cells (9-10), resembling to some extent the phenotype of cells microinjected with anti-Rac antibodies. The in vitro studies indicate that the A. proteus Rac-like protein, but not the MIHCK isoform, is engaged in the regulation of the nucleation step of the actin polymerisation process. These observations suggest that MIHCK may be one of the effectors for Rac in these extremely large cells.
机译：仍然高度了解能动的变形虫变形虫独特运动的分子机制。最近，我们已经表明，阻断内源性变形虫类Rac样蛋白会导致这些大型迁移细胞的外观发生明显且不可逆的变化，并显着抑制其运动。为了阐明变形虫变形杆菌Rac途径的机制，我们测试了抗内源性肌球蛋白I重链激酶（MIHCK）（即棘形棘轮虫和盘基网柄菌的Rac效应物之一）与抗MIHCK抗体在迁移的变形虫中的作用，以及抑制Rac和MIHCK对肌动蛋白聚合过程的影响。针对castellanii的MIHCK抗体检测到一种变形杆菌蛋白，其分子量（约95 kDa）类似于castellanii激酶。变形杆菌中MIHCK的细胞分布与变形虫中Rac-like蛋白和castellanii中的MIHCK的细胞分布非常相似。显微注射抗MIHCK抗体的变形虫运动较对照组细胞（9-10）慢，突出的假足较小（5-6），在某种程度上类似于显微注射抗Rac抗体的细胞表型。体外研究表明变形杆菌Aac Rac样蛋白参与了肌动蛋白聚合过程成核步骤的调控，但未参与MIHCK亚型。这些观察结果表明，MIHCK可能是这些极大细胞中Rac的效应子之一。

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