Oyen Mattias, Jäntti Jussi, Keränen Sirkka, Ronne Hans
Department of Plant Biology and Forest Genetics, Swedish University of Agricultural Sciences, Box 7080, 75007 Uppsala, Sweden.
Curr Genet. 2004 Feb;45(2):76-82. doi: 10.1007/s00294-003-0462-8. Epub 2003 Dec 2.
The yeast Saccharomyces cerevisiae has two closely related plasma membrane syntaxins, Sso1p and Sso2p, which together provide an essential function in vegetative cells. However, Sso1p is also specifically needed during sporulation; and this function cannot be provided by Sso2p. We used fusions between SSO1 and SSO2 to map the sporulation-specific function of SSO1. We found that the two N-terminal alpha-helices Ha and Hb of Sso1p are important for sporulation, since it is reduced 8-fold for fusions where Ha and Hb are derived from Sso2p. In contrast, the C-terminal half of Sso1p does not seem to be specifically required for sporulation. Surprisingly, we further found that the 3' untranslated region (3'UTR) of SSO1 is essential for sporulation. Western blots failed to reveal a preferential expression of Sso1p in sporulating cells, indicating that effects on gene expression are unlikely to explain why the SSO1 3'UTR is needed for sporulation.
酿酒酵母有两种密切相关的质膜 syntaxin,即 Sso1p 和 Sso2p,它们共同为营养细胞提供重要功能。然而,在孢子形成过程中也特别需要 Sso1p;而 Sso2p 无法提供此功能。我们利用 SSO1 和 SSO2 之间的融合来定位 SSO1 的孢子形成特异性功能。我们发现,Sso1p 的两个 N 端α螺旋 Ha 和 Hb 对孢子形成很重要,因为当 Ha 和 Hb 来自 Sso2p 时,融合蛋白的孢子形成能力降低了 8 倍。相比之下,Sso1p 的 C 端似乎对孢子形成并非特别必需。令人惊讶的是,我们进一步发现 SSO1 的 3'非翻译区(3'UTR)对孢子形成至关重要。蛋白质免疫印迹未能揭示 Sso1p 在孢子形成细胞中的优先表达,这表明对基因表达的影响不太可能解释为何孢子形成需要 SSO1 的 3'UTR。