The cis-Golgi Drosophila GMAP has a role in anterograde transport and Golgi organization in vivo, similar to its mammalian ortholog in tissue culture cells
The cis-Golgi Drosophila GMAP has a role in anterograde transport and Golgi organization in vivo, similar to its mammalian ortholog in tissue culture cells
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DOI：
10.1016/j.ejcb.2006.07.001
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发表时间：
2006-11-01
影响因子：
6.6
通讯作者：
Therond, Pascal
中科院分区：
文献类型：
--
作者：
Friggi-Grelin, Florence;Rabouille, Catherine;Therond, Pascal
210 kDa的高尔基体微管相关蛋白（GMAP-210）已被证明在哺乳动物组织培养细胞中高尔基体的组装和维持中发挥作用。为了建立一个遗传模型来研究GMAP-210在体内的功能，我们鉴定了它在果蝇中的近亲dGMAP。我们发现，dGMAP基因编码两个选择性剪接的转录本，其中只有一个被翻译成蛋白质产物。为了深入了解dGMAP的作用，我们产生了多克隆抗体并研究了发育过程中的蛋白质分布。该基因在胚胎和幼虫发育过程中广泛表达，在极细胞、肠道和唾液腺中表达水平最高。我们进一步表明，dGMAP是存在于高尔基体，并使用电子显微镜的唾液腺，我们观察到一个优先本地化的顺式侧和在边缘的高尔基堆栈。最后，我们证明，dGMAP在唾液腺中的过表达损害高尔基体的结构和功能，而RNAi介导的dGMAP的消耗不诱导高尔基体的任何结构修饰，并且顺行运输似乎不受影响。总之，我们的结果表明，dGMAP是哺乳动物GMAP-210的直系同源物。(c)2006年Elsevier GmbH。All rights reserved.
The Golgi microtubule-associated protein of 210 kDa (GMAP-210) has been shown to play a role in the assembly and maintenance of the Golgi apparatus in mammalian tissue culture cells. To develop a genetic model to study the function of GMAP-210 in vivo, we identified its closest relative in Drosophila melanogaster, dGMAP. We show that the dGMAP gene encodes two alternatively spliced transcripts, only one of which is translated into a protein product. To gain insight into the role of dGMAP, we generated a polyclonal antibody and investigated the protein distribution during development. This gene is ubiquitously expressed during embryonic and larval development with the highest level in polar cells, gut and salivary glands. We further show that dGMAP is present in the Golgi apparatus, and using electron microscopy of salivary glands, we observed a preferential localization at the cis side and at the rims of the Golgi stacks. Finally, we demonstrate that overexpression of dGMAP in salivary glands impairs Golgi architecture and function, whereas RNAi-mediated depletion of dGMAP does not induce any structural modification of the Golgi apparatus, and anterograde transport is seemingly unaffected. Altogether our results suggest that dGMAP is the ortholog of mammalian GMAP-210. (c) 2006 Elsevier GmbH. All rights reserved.