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Expanding the role of the dynein regulatory complex to non-axonemal functions: Association of GAS11 with the Golgi apparatus

机译：将动力蛋白调节复合物的作用扩展到非轴索功能：GAS11与高尔基体的关联

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The mammalian GAS11 gene is a candidate tumor suppressor of unknown function that was previously identified as one of several genes upregulated upon growth arrest. Interestingly, although GAS11 homologs in Trypanosoma brucei (trypanin) and Chlamydomonas reinhardtii (PF2) are integral components of the flagellar axoneme and are necessary for regulating flagellar beat, the GAS11 gene was discovered based on its expression in cells that do not assemble a motile cilium. This suggests that GAS11 function might not be restricted to the cilium. To investigate this possibility, we generated GAS11-specific antibodies and demonstrate here that GAS11 is expressed in a variety of mammalian cells that lack a motile cilium. In COS7 cells, GAS11 is associated with the detergent-insoluble cytoskeleton and exhibits a juxtanuclear localization that overlaps with the pericentrosomal Golgi apparatus. This localization is dependent upon intact microtubules and is cell-cycle regulated, such that GAS11 is dispersed throughout the cytoplasm as cells progress through mitosis. GAS11 remains associated with Golgi fragments following depolymerization of cytoplasmic microtubules but is dispersed upon disruption of the Golgi with brefeldin A. These data suggest that GAS11 is associated with the Golgi apparatus. In support of this, recombinant GAS11 binds Golgi membranes in vitro. In growth-arrested mIMCD3 cells, GAS11 co-localizes with gamma-tubulin at the base of the primary cilium. The pericentrosomal Golgi apparatus and base of the cilium both represent convergence points for microtubule minus ends and correspond to sites where dynein regulation is required. The algal GAS11 homolog functions as part of a dynein regulatory complex (DRC) in the axoneme (Rupp and Porter. J Cell Biol 2003;162:47-57) and our findings suggest that components of this axonemal dynein regulatory system have been adapted in mammalian cells to participate in non-axonemal functions.
机译：哺乳动物GAS11基因是未知功能的候选肿瘤抑制基因，先前被鉴定为生长停滞后上调的几种基因之一。有趣的是，尽管布鲁氏锥虫（trypanin）和莱茵衣藻（PF2）中的GAS11同源物是鞭毛轴突的组成部分，并且是调节鞭毛搏动所必需的，但GAS11基因是根据其在不装配能动纤毛的细胞中的表达而发现的。这表明GAS11功能可能不限于纤毛。为了研究这种可能性，我们生成了GAS11特异性抗体，并在这里证明GAS11在缺乏运动纤毛的多种哺乳动物细胞中表达。在COS7细胞中，GAS11与去污剂不溶的细胞骨架相关，并表现出与中心周高尔基体重叠的近核定位。该定位取决于完整的微管，并且受细胞周期调节，从而当细胞通过有丝分裂进行时，GAS11分散在整个细胞质中。在胞质微管解聚后，GAS11仍与高尔基体片段相关，但在布雷菲德菌素A破坏高尔基体后，GAS11分散。这些数据表明GAS11与高尔基体相关。为此，重组GAS11在体外与高尔基体膜结合。在生长停滞的mIMCD3细胞中，GAS11与γ-微管蛋白共定位在初级纤毛的基部。中心体高尔基体和纤毛的基部都代表微管负端的会聚点，并且对应于需要动力蛋白调节的部位。藻类GAS11同源物在轴索蛋白中充当动力蛋白调节复合物（DRC）的一部分（Rupp and Porter。J Cell Biol 2003; 162：47-57），我们的发现表明，该轴索动力蛋白调节系统的成分已在哺乳动物细胞参与非轴突功能。

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