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拟南芥DOK1编码一种参与生殖过程的功能性多萜醇激酶。

Arabidopsis DOK1 encodes a functional dolichol kinase involved in reproduction.

作者信息

Kanehara Kazue, Cho Yueh, Lin Ying-Chen, Chen Chia-En, Yu Chao-Yuan, Nakamura Yuki

机构信息

Academia Sinica, Institute of Plant and Microbial Biology, No. 128 Sec. 2 Academia Rd., Nankang Taipei, 11529, Taiwan; Molecular and Biological Agricultural Sciences Program, Academia Sinica, Taiwan International Graduate Program, No. 128 Sec. 2 Academia Rd., Nankang, Taipei, 11529, Taiwan; Biotechnology Center, National Chung-Hsing University, 250 Kuo-Kuang Rd., Taichung, 402, Taiwan; Department of Applied Science and Engineering, Muroran Institute of Technology, 27-1 Mizumoto-cho, Muroran, Hokkaido, 050-8585, Japan.

出版信息

Plant J. 2015 Jan;81(2):292-303. doi: 10.1111/tpj.12727. Epub 2014 Dec 16.

Abstract

Dolichol phosphate (Dol-P) serves as a carrier of complex polysaccharides during protein glycosylation. Dol-P is synthesized by the phosphorylation of dolichol or the monodephosphorylation of dolichol pyrophosphate (Dol-PP); however, the enzymes that catalyze these reactions remain unidentified in Arabidopsis thaliana. We performed a genome-wide search for cytidylyltransferase motif-containing proteins in Arabidopsis, and found that At3g45040 encodes a protein homologous with Sec59p, a dolichol kinase (DOK) in Saccharomyces cerevisiae. At3g45040, designated AtDOK1, complemented defects in the growth and N-linked glycosylation of the S. cerevisiae sec59 mutant, suggesting that AtDOK1 encodes a functional DOK. To characterize the physiological roles of AtDOK1 in planta, we isolated two independent lines of T-DNA-tagged AtDOK1 mutants, dok1-1 and dok1-2. The heterozygous plants showed developmental defects in male and female gametophytes, including an aberrant pollen structure, low pollen viability, and short siliques. Additionally, the mutations had incomplete penetrance. These results suggest that AtDOK1 is a functional DOK required for reproductive processes in Arabidopsis.

摘要

多萜醇磷酸酯(Dol-P)在蛋白质糖基化过程中作为复合多糖的载体。Dol-P通过多萜醇的磷酸化或多萜醇焦磷酸酯(Dol-PP)的单去磷酸化合成;然而,在拟南芥中催化这些反应的酶仍未确定。我们在拟南芥中对含胞苷转移酶基序的蛋白质进行了全基因组搜索,发现At3g45040编码一种与酿酒酵母中的多萜醇激酶(DOK)Sec59p同源的蛋白质。At3g45040被命名为AtDOK1,它弥补了酿酒酵母sec59突变体生长和N-连接糖基化的缺陷,这表明AtDOK1编码一种功能性DOK。为了表征AtDOK1在植物中的生理作用,我们分离了两个独立的T-DNA标签AtDOK1突变体系,dok1-1和dok1-2。杂合植物在雄配子体和雌配子体中表现出发育缺陷,包括异常的花粉结构、低花粉活力和短角果。此外,这些突变具有不完全的外显率。这些结果表明AtDOK1是拟南芥生殖过程所需的功能性DOK。

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