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Functional analysis of GS28, an intra-Golgi SNARE, in Caenorhabditis elegans

机译：秀丽隐杆线虫GS28（高尔基体内SNARE）的功能分析

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摘要

Intra-Golgi retrograde transport is assumed to maintain Golgi function by recycling Golgi-resident proteins to younger cisternae in the progression of entire Golgi stack from cis to trans. GS28 (Golgi SNARE of 28 kDa, also known as GOS28) is a Golgi-localized SNARE protein and has been implicated in intra-Golgi retrograde transport. However, the in vivo functions of GS28, and consequently, the roles of the intra-Golgi retrograde transport in animal development are largely unknown. In this study, we generated deletion mutants of Caenorhabditis elegans GS28 and performed a synthetic lethal RNAi screen using GS28 mutants. We found that another Golgi-localized SNARE, Ykt6, functions cooperatively with GS28 in embryonic development. During post-embryonic development, GS28 mutants exhibited reduced seam cell numbers and a missing ray phenotype under Ykt6 knockdown conditions, suggesting that cell proliferation and/or differentiation of stem cell-like seam cells are impaired in GS28- and Ykt6-depleted worms. We also demonstrated that GS28 and Ykt6 act redundantly for the proper expression of Golgi-resident proteins in adult intestinal cells. This study reveals the in vivo importance of the Golgi-localized SNAREs GS28 and Ykt6.
机译：高尔基体内逆行转运被认为通过在整个高尔基体从顺式到反式的过程中将高尔基体驻留蛋白循环到年轻的池中来维持高尔基体功能。 GS28（28 kDa的高尔基SNARE，也称为GOS28）是高尔基体定位的SNARE蛋白，与高尔基体内逆行运输有关。然而，GS28的体内功能，因此，高尔基体内逆行运输在动物发育中的作用尚不清楚。在这项研究中，我们生成了秀丽隐杆线虫GS28的缺失突变体，并使用GS28突变体进行了合成的致死性RNAi筛选。我们发现另一个高尔基体定位的SNARE，Ykt6，在胚胎发育中与GS28协同作用。在胚后发育过程中，GS28突变体在Ykt6敲低条件下显示出减少的接缝细胞数量和缺失的射线表型，表明在GS28和Ykt6缺失的蠕虫中，细胞增殖和/或干细胞样接缝细胞的分化受到损害。我们还证明了GS28和Ykt6对于在成年肠道细胞中高尔基体驻留蛋白的正确表达有多余的作用。这项研究揭示了高尔基体定位的SNARE GS28和Ykt6在体内的重要性。

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