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玉米具短尖突变是16 kDa γ-玉米醇溶蛋白基因中的一个缺失,它会引发未折叠蛋白反应。

The maize Mucronate mutation is a deletion in the 16-kDa gamma-zein gene that induces the unfolded protein response.

作者信息

Kim Cheol Soo, Gibbon Bryan C, Gillikin Jeffrey W, Larkins Brian A, Boston Rebecca S, Jung Rudolf

机构信息

Department of Plant Sciences, University of Arizona, Tucson, AZ 85721, USA.

出版信息

Plant J. 2006 Nov;48(3):440-51. doi: 10.1111/j.1365-313X.2006.02884.x. Epub 2006 Sep 29.

Abstract

Mucronate (Mc) was identified as a dominant maize (Zea mays L.) opaque kernel mutation that alters zein storage protein synthesis. Zein protein bodies in Mc endosperm are misshapen and are associated with increased levels of ER Lumenal Binding Protein (BiP). Using GeneCalling to profile endosperm RNA transcripts, we identified an aberrant RNA in Mc that encodes the 16-kDa gamma-zein protein. The transcript contains a 38-bp deletion (nucleotides 406-444 after the initiation codon) that creates a frame-shift mutation and an abnormal sequence for the last 63 amino acids. Genetic mapping revealed the Mc mutation is linked with the locus encoding the 16-kDa gamma-zein, and two-dimensional gel electrophoresis confirmed the 16-kDa gamma-zein protein is altered in Mc. The mutant protein exhibited changes in solubility properties and co-immunoprecipitated with the molecular chaperone, BiP. Transgenic maize plants expressing the Mc 16-kDa gamma-zein manifested an opaque kernel phenotype with enhanced levels of BiP in the endosperm, similar to the Mc mutant. Unlike the wild-type protein, the Mc 16-kDa gamma-zein interacted only weakly with the 22-kDa alpha-zein when expressed in the yeast two-hybrid system. These results indicate that the Mc phenotype results from a frame-shift mutation in the gene encoding the 16-kDa gamma-zein protein, leading to the unfolded protein response in developing endosperm.

摘要

具短尖的(Mc)被鉴定为一种显性玉米(Zea mays L.)不透明籽粒突变体,它会改变醇溶蛋白储存蛋白的合成。Mc胚乳中的醇溶蛋白体形状异常,并与内质网腔结合蛋白(BiP)水平升高有关。利用基因表达谱分析胚乳RNA转录本,我们在Mc中鉴定出一种异常RNA,它编码16 kDa的γ-醇溶蛋白。该转录本包含一个38 bp的缺失(起始密码子后第406 - 444个核苷酸),导致移码突变以及最后63个氨基酸的异常序列。遗传图谱显示Mc突变与编码16 kDaγ-醇溶蛋白的基因座连锁,二维凝胶电泳证实Mc中16 kDaγ-醇溶蛋白发生了改变。突变蛋白的溶解性发生了变化,并与分子伴侣BiP进行了共免疫沉淀。表达Mc 16 kDaγ-醇溶蛋白的转基因玉米植株表现出不透明籽粒表型,胚乳中BiP水平升高,类似于Mc突变体。与野生型蛋白不同,Mc 16 kDaγ-醇溶蛋白在酵母双杂交系统中表达时,与22 kDaα-醇溶蛋白的相互作用较弱。这些结果表明,Mc表型是由编码16 kDaγ-醇溶蛋白的基因发生移码突变导致的,从而在发育中的胚乳中引发了未折叠蛋白反应。

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