Center for Plant Molecular Biology (ZMBP), University of Tübingen, 72076 Tübingen, Germany.
Center for Plant Molecular Biology (ZMBP), University of Tübingen, 72076 Tübingen, Germany
Plant Physiol. 2018 Apr;176(4):2886-2903. doi: 10.1104/pp.17.01260. Epub 2018 Mar 1.
Alternative splicing (AS) is prevalent in higher eukaryotes, and generation of different AS variants is tightly regulated. Widespread AS occurs in response to altered light conditions and plays a critical role in seedling photomorphogenesis, but despite its frequency and effect on plant development, the functional role of AS remains unknown for most splicing variants. Here, we characterized the light-dependent AS variants of the gene encoding the splicing regulator Ser/Arg-rich protein SR30 in Arabidopsis (). We demonstrated that the splicing variant , which is predominantly produced in darkness, is enriched within the nucleus and strongly depleted from ribosomes. Light-induced AS from a downstream 3' splice site gives rise to , which is exported to the cytosol and translated, coinciding with SR30 protein accumulation upon seedling illumination. Constitutive expression of SR30.1 and SR30.2 fused to fluorescent proteins revealed their identical subcellular localization in the nucleoplasm and nuclear speckles. Furthermore, expression of either variant shifted splicing of a genomic reporter toward , suggesting that an autoregulatory feedback loop affects splicing. We provide evidence that can be further spliced and, unlike , the resulting cassette exon variant is sensitive to nonsense-mediated decay. Our work delivers insight into the complex and compartmentalized RNA processing mechanisms that control the expression of the splicing regulator SR30 in a light-dependent manner.
可变剪接(AS)在高等真核生物中很普遍,不同 AS 变体的产生受到严格调控。广泛的 AS 是对光照条件改变的响应,在幼苗光形态建成中起着关键作用,但尽管 AS 的频率和对植物发育的影响很大,但大多数剪接变体的功能作用仍然未知。在这里,我们对拟南芥(Arabidopsis)中编码剪接调节因子丝氨酸/精氨酸丰富蛋白 SR30 的基因的光依赖性 AS 变体进行了表征()。我们证明了主要在黑暗中产生的剪接变体 ,在核内富集,而在核糖体中强烈缺失。来自下游 3' 剪接位点的光诱导 AS 产生 ,该变体被输出到细胞质并翻译,这与幼苗光照时 SR30 蛋白的积累相吻合。荧光蛋白融合的 SR30.1 和 SR30.2 的组成型表达揭示了它们在核质和核斑中的相同亚细胞定位。此外,任一变体的表达都将基因组 报告基因的剪接偏向 ,表明一个自反馈回路影响 剪接。我们提供的证据表明 可以进一步剪接,与 不同的是,由此产生的外显子剪接变体 对无意义介导的衰变敏感。我们的工作深入了解了复杂的、区室化的 RNA 处理机制,这些机制以光照依赖的方式控制剪接调节因子 SR30 的表达。