Fomonov G V, Ter-Avanesian M D
Mol Biol (Mosk). 2005 Sep-Oct;39(5):859-68.
The Mcd4 protein of Saccharomyces cerevisiae is probably involved in addition of the phosphoethanolamine moiety to the first mannose residue of the glycosylphosphatidylinositol precursor(s). However, significance of this modification is unclear. Besides, functions of the MCD4 gene also is not completely clear, since mutations in this gene may have pleiotropic manifestations, which are not obviously related to the glycosylphosphatidylinositol biosynthesis. To clarify the functions of Mcd4p we have performed a search for genes whose mutations are lethal or semilethal in combination with the ssu21 mutation in MCD4. In total, we have isolated six mutations some of which cause sensitivity to SDS and/or calcofluor white. Genes which are able to complement two of these mutations were cloned. They were MNN9 which encodes protein involved in formation of outer chains of the N-linked glycans of secretory proteins and GWT1, encoding the protein of the endoplasmic reticulum involved in the glycosylphosphatidylinositol biosynthesis. The results obtained indicate that in both cases growth inhibition was caused by defect of cell wall biogenesis and alteration of folding of secretory proteins. Search for mutations that lethal in combination with the ssu21 is an effective approach to reveal genes involved in the control of cell wall biogenesis.
酿酒酵母的Mcd4蛋白可能参与将磷酸乙醇胺部分添加到糖基磷脂酰肌醇前体的第一个甘露糖残基上。然而,这种修饰的意义尚不清楚。此外,MCD4基因的功能也不完全清楚,因为该基因的突变可能具有多效性表现,而这些表现与糖基磷脂酰肌醇生物合成并无明显关联。为了阐明Mcd4p的功能,我们对那些与MCD4中的ssu21突变相结合时其突变具有致死性或半致死性的基因进行了搜索。总共,我们分离出了六个突变,其中一些会导致对SDS和/或荧光增白剂的敏感性。能够互补其中两个突变的基因被克隆出来。它们是MNN9,其编码参与分泌蛋白N - 连接聚糖外链形成的蛋白质,以及GWT1,其编码参与糖基磷脂酰肌醇生物合成的内质网蛋白。所获得的结果表明,在这两种情况下,生长抑制都是由细胞壁生物合成缺陷和分泌蛋白折叠改变引起的。搜索与ssu21相结合时具有致死性的突变是揭示参与细胞壁生物合成控制的基因的有效方法。