Role of the Small GTPase Rho3 in Golgi/Endosome trafficking through functional interaction with adaptin in Fission Yeast.
Role of the Small GTPase Rho3 in Golgi/Endosome trafficking through functional interaction with adaptin in Fission Yeast.
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DOI：
10.1371/journal.pone.0016842
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发表时间：
2011-02-03
期刊：
影响因子：
3.7
通讯作者：
Sugiura R
中科院分区：
文献类型：
--
作者：
Kita A;Li C;Yu Y;Umeda N;Doi A;Yasuda M;Ishiwata S;Taga A;Horiuchi Y;Sugiura R
我们之前已经鉴定了apm 1+的突变等位基因，该突变等位基因编码网格蛋白相关衔接蛋白-1（AP-1）复合物的哺乳动物µ1A亚基的同源物，并且我们证明了Apm 1在裂殖酵母中高尔基体/内体运输、分泌和液泡融合中的作用。在本研究中，我们分离的rho 3+，它编码一个Rho家族的小GTdR，一个重要的调节外囊形成，作为apm 1 -1突变细胞的温度敏感性生长的多拷贝抑制剂。Rho 3的过表达抑制apm 1 -1突变细胞的Cl−敏感性和免疫抑制剂敏感性。Rho 3的过表达还抑制了空泡的破碎，以及v-SNARE Syb 1在高尔基体/内体中的积累，并部分抑制了与apm 1缺失细胞相关的分泌缺陷。值得注意的是，电子显微镜观察的rho 3缺失细胞显示异常高尔基体样结构的积累，空泡破碎，和分泌囊泡的积累，这些表型是非常相似的apm 1缺失细胞。此外，rho 3缺失细胞和apm 1缺失细胞表现出非常相似的表型特征，包括对免疫抑制剂FK 506、细胞壁损伤剂米卡芬净、Cl-和丙戊酸的敏感性。绿色荧光蛋白（GFP）-Rho 3定位于高尔基体/内体以及质膜和分裂位点。最后，Rho 3被证明与Apm 1以及网格蛋白相关的AP-1复合物的其他亚基以GTP和效应结构域依赖的方式形成复合物。两者合计，我们的研究结果揭示了一个新的作用Rho 3在调节高尔基体/内体贩运，并建议网格蛋白相关的衔接蛋白-1和Rho 3协调在细胞内运输的裂殖酵母。据我们所知，这项研究提供了第一个证据的直接联系之间的小GTdR Rho和网格蛋白相关的衔接蛋白-1膜贩运。
We had previously identified the mutant allele of apm1+ that encodes a homolog of the mammalian µ1A subunit of the clathrin-associated adaptor protein-1 (AP-1) complex, and we demonstrated the role of Apm1 in Golgi/endosome trafficking, secretion, and vacuole fusion in fission yeast. In the present study, we isolated rho3+, which encodes a Rho-family small GTPase, an important regulator of exocystosis, as a multicopy-suppressor of the temperature-sensitive growth of the apm1-1 mutant cells. Overexpression of Rho3 suppressed the Cl− sensitivity and immunosuppressant sensitivity of the apm1-1 mutant cells. Overexpression of Rho3 also suppressed the fragmentation of vacuoles, and the accumulation of v-SNARE Syb1 in Golgi/endosomes and partially suppressed the defective secretion associated with apm1-deletion cells. Notably, electron microscopic observation of the rho3-deletion cells revealed the accumulation of abnormal Golgi-like structures, vacuole fragmentation, and accumulation of secretory vesicles; these phenotypes were very similar to those of the apm1-deletion cells. Furthermore, the rho3-deletion cells and apm1-deletion cells showed very similar phenotypic characteristics, including the sensitivity to the immunosuppressant FK506, the cell wall-damaging agent micafungin, Cl−, and valproic acid. Green fluorescent protein (GFP)-Rho3 was localized at Golgi/endosomes as well as the plasma membrane and division site. Finally, Rho3 was shown to form a complex with Apm1 as well as with other subunits of the clathrin-associated AP-1 complex in a GTP- and effector domain-dependent manner. Taken together, our findings reveal a novel role of Rho3 in the regulation of Golgi/endosome trafficking and suggest that clathrin-associated adaptor protein-1 and Rho3 co-ordinate in intracellular transport in fission yeast. To the best of our knowledge, this study provides the first evidence of a direct link between the small GTPase Rho and the clathrin-associated adaptor protein-1 in membrane trafficking.
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影响因子：
9.2
作者：
Bone, N;Millar, JBA;Armstrong, J
通讯作者：
Armstrong, J
影响因子：
4
作者：
Nakano, K;Imai, J;Mabuchi, I
通讯作者：
Mabuchi, I
影响因子：
3.3
作者：
Wu H;Turner C;Gardner J;Temple B;Brennwald P
通讯作者：
Brennwald P
影响因子：
64.5
作者：
NOVICK, P;FERRO, S;SCHEKMAN, R
通讯作者：
SCHEKMAN, R
DOI：
10.1016/j.jpba.2010.07.009
发表时间：
2010-12-15
影响因子：
3.4
作者：
Taga, Atsushi;Satoh, Ryosuke;Sugiura, Reiko
通讯作者：
Sugiura, Reiko