Related to ubiquitin 1 and 2 are redundant and essential and regulate vegetative growth, auxin signaling, and ethylene production in Arabidopsis
Related to ubiquitin 1 and 2 are redundant and essential and regulate vegetative growth, auxin signaling, and ethylene production in Arabidopsis
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DOI：
10.1105/tpc.104.024943
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发表时间：
2004-09-01
期刊：
影响因子：
11.6
通讯作者：
Callis, J
中科院分区：
文献类型：
--
作者：
Bostick, M;Lochhead, SR;Callis, J
与泛素(RUB)/Nedd8相关的是一种泛素样蛋白，共价附着在cullins上，cullins是SCF （Skp， Cdc53p/Cul1和F-box蛋白）复合物的一个亚基，是E3泛素连接酶，并且已被证明是该复合物强大功能所必需的。以拟南芥为研究对象，研究了RUB1和RUB2两种RUB1和RUB2蛋白水平降低的影响。对单个rub编码位点纯合的T-DNA插入空系进行了分析，发现其具有野生型表型。双突变体从未恢复。在具有单一功能的rubb编码基因的植物中，超过四分之一的自交受精后代在两细胞阶段的胚胎中死亡。异交表明，该空等位基因从父本和母本的遗传都减少了。血红素标记的RUB1和RUB2结合到相同的cullin蛋白CUL1上，并产生相同的结合模式。为了进一步了解RUB1蛋白的功能，我们将一个设计用于产生双链RUB1 mRNA的构建体引入植物，并对三个RUB1-和rub2编码mRNA和RUB1/2蛋白含量降低的品系进行了详细的分析。成熟植株严重矮化，幼苗在根系试验中对生长素不敏感，深色生长的幼苗具有部分三重反应表型，当幼苗生长在乙烯感知或合成抑制剂上时，这种表型被抑制。灌丛系的乙烯产量是野生型的三到五倍。本研究表明RUB1和RUB2在遗传和生化上都是冗余的，并表明RUB1/2蛋白对早期胚胎细胞分裂至关重要，它们调节着多种过程。
Related to Ubiquitin (RUB)/Nedd8 is a ubiquitin-like protein that covalently attaches to cullins, a subunit of the SCF (for Skp, Cdc53p/Cul1, and F-box protein) complex, an E3 ubiquitin ligase, and has been shown to be required for robust function of the complex. The effects of reducing protein levels for two Rub proteins, RUB1 and RUB2, were characterized in Arabidopsis thaliana. T-DNA insertional null lines homozygous at a single RUB-encoding locus were analyzed and found to have a wild-type phenotype. A double mutant was never recovered. More than one-quarter of the progeny from the self-fertilization of plants with a single functional RUB-encoding gene died as embryos at the two-cell stage. Outcrosses demonstrated reduced inheritance of the null allele from both the male and female parent. Hemigglutinin-tagged forms of RUB1 and RUB2 conjugate to the same cullin protein, CUL1, and produce the same conjugation pattern. To further understand the function of the RUB proteins, a construct designed to produce a double-stranded RUB1 mRNA was introduced into plants, and three lines with reduced levels of RUB1- and RUB2-encoding mRNA and RUB1/2 protein content were analyzed in detail. Mature plants were severely dwarfed, seedlings were insensitive to auxin in root assays, and dark-grown seedlings had a partial triple-response phenotype that was suppressed when seedlings were grown on ethylene perception or synthesis inhibitors. The dsrub lines produced threefold to fivefold more ethylene than the wild type. This study illustrates that RUB1 and RUB2 are genetically and biochemically redundant and demonstrates that RUB1/2 proteins are essential for early embryonic cell divisions and that they regulate diverse processes.