Molecular cloning and characterization of a Pichia pastoris ortholog of the yeast Golgi GDP-mannose transporter gene
Molecular cloning and characterization of a Pichia pastoris ortholog of the yeast Golgi GDP-mannose transporter gene
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DOI：
10.2323/jgam.52.137
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发表时间：
2006-06-01
影响因子：
1.2
通讯作者：
Yoda, Koji
中科院分区：
文献类型：
--
作者：
Arakawa, Kumiko;Abe, Masato;Yoda, Koji
酵母高尔基体有两种结构特征。酿酒酵母的高尔基体由许多分散在细胞质中的囊泡区室组成，这些区室被大量高尔基体标记蛋白识别。相比之下，通过电子显微镜和一些标记蛋白的免疫荧光染色，毕赤酵母的高尔基体被组织在位于移行内质网（tER）位点附近的少量堆叠池中。二磷酸鸟苷 (GDP)-甘露糖转运蛋白 (GMT) 是酵母高尔基体的重要组成部分。我们分离了毕赤酵母 GMT 基因的直系同源物，并通过表位标记可视化基因产物，以验证高尔基体的结构特征。在巴斯德毕赤酵母细胞中的标记产物在杆状区室中观察到，其中还发现了 Och1 甘露糖基转移酶，并且 tER 标记 Sec12 和 Sec13 蛋白位于非常靠近它们的位置。目前的结果进一步证明了毕赤酵母细胞中高尔基体的有限定位。
There are two structural profiles in the yeast Golgi. The Golgi of Saccharomyces cerevisiae is composed of a number of vesicular compartments dispersed in the cytoplasm as recognized by a large number of Golgi marker proteins. In contrast, the Golgi of Pichia pastoris was reported to be organized in a small number of stacked cisternae located near the transitional endoplasmic reticulum (tER) sites by electron microscopy and immunofluorescent staining of a few marker proteins. The guanosine diphosphate (GDP)-mannose transporter (GMT) is an essential component in the yeast Golgi apparatus. We isolated an ortholog of the GMT gene of P. pastoris and visualized the gene product by epitope tagging to verify the structural characteristics of the Golgi. The tagged product in P. pastoris cell was observed in rod-like compartments in which Och1 mannosyltransferase was also found and the tER marker Sec12 and Sec13 proteins localized very close to them. The present results add further evidence of the restricted localization of the Golgi in P. pastoris cell.