Yeast functional analysis:: Identification of two essential genes involved in ER to Golgi trafficking
Yeast functional analysis:: Identification of two essential genes involved in ER to Golgi trafficking
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DOI：
10.1034/j.1600-0854.2003.00116.x
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发表时间：
2003-09-01
期刊：
影响因子：
4.5
通讯作者：
Haguenauer-Tsapis, R
中科院分区：
文献类型：
--
作者：
Belgareh-Touzé, N;Corral-Debrinski, M;Haguenauer-Tsapis, R
我们筛选了61个酵母菌株的潜在涉及分泌和空泡途径的基因，每个酵母菌株携带一个基本的孤儿基因，该基因受Teto(7)-Cyc1启动子调控，可被多西环素下调。在下调这些基因的表达后，我们对分泌和空泡途径的标志物进行了系统的Western印迹分析，这些标志物在细胞内运输时经历了翻译后的修饰。蛋白质前体的积累，通过免疫印迹分析，表明分泌途径或相关的生化修饰存在缺陷。在对整个文库进行筛选后，我们鉴定了两个参与内质网到高尔基体运输的基因：顺式-戊烯基转移酶RER2和功能未知的USE1。我们证明，抑制USE1也导致BIP的分泌，因此除了影响顺行的ER到高尔基体的运输外，还可能影响逆行。该集合还包括参与细胞内运输的两个必要基因，它们被方便地抑制，而不会导致生长或运输缺陷。
We screened for genes potentially involved in the secretory and vacuolar pathways a collection of 61 yeast strains, each bearing an essential orphan gene regulated by the tetO(7) -CYC1 promoter that can be down-regulated by doxycycline. After down-regulating the expression of these genes, we performed systematic Western blot analysis for markers of the secretory and vacuolar pathways that undergo post-translational modifications in their intracellular trafficking. Accumulation of protein precursors, revealed by Western immunoblot analysis, indicates defects in the secretory pathway or in associated biochemical modifications. After screening the whole collection, we identified two genes involved in ER to Golgi trafficking: RER2 , a cis -prenyl transferase, and USE1 , the function of which was unknown. We demonstrated that repression of USE1 also leads to BiP secretion, and therefore likely affects retrograde, in addition to anterograde, ER to Golgi trafficking. The collection also includes two essential genes involved in intracellular trafficking that were conveniently repressed without resulting growth or trafficking defects.