Identification of the region of rice 13 kDa prolamin essential for the formation of ER-derived protein bodies using a heterologous expression system.
Identification of the region of rice 13 kDa prolamin essential for the formation of ER-derived protein bodies using a heterologous expression system.
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使用异源表达系统鉴定水稻 13 kDa 谷醇溶蛋白区域对于形成 ER 衍生蛋白体至关重要。
DOI：
10.1080/09168451.2014.991684
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Shigeru Satoh
中科院分区：
文献类型：
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作者：
Takehiro Masumura**;Takanari Shigemitsu**;Shigeto Morita;Shigeru Satoh
谷类醇溶蛋白是一种醇溶种子贮藏蛋白，可在异源组织中诱导内质网衍生蛋白小体(PBS)。与玉米和小麦醇溶蛋白一样，水稻醇溶蛋白可以形成内质网衍生的PBS，但对PB形成至关重要的成熟多肽区域尚未确定。在这项研究中，我们通过在酵母等异源组织中表达融合了绿色荧光蛋白(GFP)的13 kDa醇溶蛋白缺失突变体来研究内质网衍生的PB样结构的形成机制。13 kDa醇溶蛋白-GFP融合蛋白在转基因酵母中稳定积累，并形成内质网衍生的PB样结构。相反，水稻α-球蛋白-绿色荧光蛋白融合蛋白被运输到液泡中。此外，13 kDa醇溶蛋白的中间和COOH末端区域形成内质网衍生的PB样结构，而13 kDa醇溶蛋白的NH2末端区域不形成这种结构。这些结果表明，13 kDa醇溶蛋白的中间和COOH末端区域可以被保留，从而可以在酵母中诱导内质网衍生的PB。
Cereal prolamins, which are alcohol-soluble seed storage proteins, can induce ER-derived protein bodies (PBs) in heterologous tissue. Like maize and wheat prolamins, rice prolamins can form ER-derived PBs, but the region of mature polypeptides that is essential for PB formation has not been identified. In this study, we examined the formation mechanisms of ER-derived PB-like structures by expressing rice 13 kDa prolamin-deletion mutants fused to green fluorescent protein (GFP) in heterologous tissues such as yeast. The 13 kDa prolamin–GFP fusion protein was stably accumulated in transgenic yeast and formed an ER-derived PB-like structure. In contrast, rice α-globulin–GFP fusion protein was transported to vacuoles. In addition, the middle and COOH-terminal regions of 13 kDa prolamin formed ER-derived PB-like structures, whereas the NH2-terminal region of 13 kDa prolamin did not form such structures. These results suggest that the middle and COOH-terminal regions of 13 kDa prolamin can be retained and thus can induce ER-derived PB in yeast.