Golovkin Maxim, Reddy Anireddy S N
Department of Biology, Colorado State University, Fort Collins, Colorado 80523, USA.
Plant Physiol. 2003 Aug;132(4):1884-91. doi: 10.1104/pp.103.023192.
U1 small nuclear ribonucleoprotein (snRNP)-70K (U1-70K), a U1 snRNP-specific protein, is involved in the early stages of spliceosome formation. In non-plant systems, it is involved in constitutive and alternative splicing. It has been shown that U1snRNP is dispensable for in vitro splicing of some animal pre-mRNAs, and inactivation of U1-70K in yeast (Saccharomyces cerevisiae) is not lethal. As in yeast and humans (Homo sapiens), plant U1-70K is coded by a single gene. In this study, we blocked the expression of Arabidopsis U1-70K in petals and stamens by expressing U1-70K antisense transcript using the AP3 (APETALA3) promoter specific to these floral organs. Flowers of transgenic Arabidopsis plants expressing U1-70K antisense transcript showed partially developed stamens and petals that are arrested at different stages of development. In some transgenic lines, flowers have rudimentary petals and stamens and are male sterile. The severity of the phenotype is correlated with the level of the antisense transcript. Molecular analysis of transgenic plants has confirmed that the observed phenotype is not due to disruption of whorl-specific homeotic genes, AP3 or PISTILLATA, responsible for petal and stamen development. The AP3 transcript was not detected in transgenic flowers with severe phenotype. Flowers of Arabidopsis plants transformed with a reporter gene driven by the same promoter showed no abnormalities. These results show that U1-70K is necessary for the development of sepals and petals and is an essential gene in plants.
U1小核核糖核蛋白(snRNP)-70K(U1-70K)是一种U1 snRNP特异性蛋白,参与剪接体形成的早期阶段。在非植物系统中,它参与组成型和可变剪接。研究表明,U1 snRNP对于某些动物前体mRNA的体外剪接并非必需,并且酵母(酿酒酵母)中U1-70K的失活并不致命。与酵母和人类(智人)一样,植物U1-70K由单个基因编码。在本研究中,我们使用这些花器官特异的AP3(APETALA3)启动子表达U1-70K反义转录本,从而阻断拟南芥花瓣和雄蕊中U1-70K的表达。表达U1-70K反义转录本的转基因拟南芥植株的花显示出部分发育的雄蕊和花瓣,它们在不同的发育阶段停滞。在一些转基因株系中,花具有发育不全的花瓣和雄蕊,并且雄性不育。表型的严重程度与反义转录本的水平相关。转基因植物的分子分析证实,观察到的表型不是由于负责花瓣和雄蕊发育的轮特异性同源异型基因AP3或PISTILLATA的破坏所致。在具有严重表型的转基因花中未检测到AP3转录本。用相同启动子驱动的报告基因转化的拟南芥植株的花没有异常。这些结果表明,U1-70K对于萼片和花瓣的发育是必需的,并且是植物中的一个必需基因。