Du Y, Pypaert M, Novick P, Ferro-Novick S
Department of Cell Biology, Yale University, New Haven, Connecticut 06519-1418, USA.
Mol Biol Cell. 2001 Sep;12(9):2614-28. doi: 10.1091/mbc.12.9.2614.
In the yeast Saccharomyces cerevisiae, the endoplasmic reticulum (ER) is found at the periphery of the cell and around the nucleus. The segregation of ER through the mother-bud neck may occur by more than one mechanism because perinuclear, but not peripheral ER, requires microtubules for this event. To identify genes whose products are required for cortical ER inheritance, we have used a Tn3-based transposon library to mutagenize cells expressing a green fluorescent protein-tagged ER marker protein (Hmg1p). This approach has revealed that AUX1/SWA2 plays a role in ER inheritance. The COOH terminus of Aux1p/Swa2p contains a J-domain that is highly related to the J-domain of auxilin, which stimulates the uncoating of clathrin-coated vesicles. Deletion of the J-domain of Aux1p/Swa2p leads to vacuole fragmentation and membrane accumulation but does not affect the migration of peripheral ER into daughter cells. These findings suggest that Aux1p/Swa2p may be a bifunctional protein with roles in membrane traffic and cortical ER inheritance. In support of this hypothesis, we find that Aux1p/Swa2p localizes to ER membranes.
在酿酒酵母中,内质网(ER)位于细胞周边及细胞核周围。内质网通过母细胞-芽颈的分离可能通过多种机制发生,因为核周内质网而非周边内质网在此过程中需要微管。为了鉴定其产物对于皮质内质网遗传所必需的基因,我们使用了基于Tn3的转座子文库来诱变表达绿色荧光蛋白标记的内质网标记蛋白(Hmg1p)的细胞。该方法揭示了AUX1/SWA2在内质网遗传中发挥作用。Aux1p/Swa2p的COOH末端包含一个J结构域,该结构域与辅助蛋白的J结构域高度相关,辅助蛋白可刺激网格蛋白包被小泡的去包被。删除Aux1p/Swa2p的J结构域会导致液泡碎片化和膜积累,但不影响周边内质网向子细胞的迁移。这些发现表明Aux1p/Swa2p可能是一种在膜运输和皮质内质网遗传中发挥作用的双功能蛋白。为支持这一假设,我们发现Aux1p/Swa2p定位于内质网膜。