Centro de Investigación Príncipe Felipe, Gene Expression coupled to RNA Transport Laboratory, Av Saler 16. E-46012, Valencia, Spain.
Nucleic Acids Res. 2011 Oct;39(19):8599-611. doi: 10.1093/nar/gkr496. Epub 2011 Jul 12.
Efficient coupling between mRNA synthesis and export is essential for gene expression. Sus1/ENY2, a component of the SAGA and TREX-2 complexes, is involved in both transcription and mRNA export. While most yeast genes lack introns, we previously reported that yeast SUS1 bears two. Here we show that this feature is evolutionarily conserved and critical for Sus1 function. We determine that while SUS1 splicing is inefficient, it responds to cellular conditions, and intronic mutations either promoting or blocking splicing lead to defects in mRNA export and cell growth. Consistent with this, we find that an intron-less SUS1 only partially rescues sus1Δ phenotypes. Remarkably, splicing of each SUS1 intron is also affected by the presence of the other and by SUS1 exonic sequences. Moreover, by following SUS1 RNA and protein levels we establish that nonsense-mediated decay (NMD) pathway and the splicing factor Mud2 both play a role in SUS1 expression. Our data (and those of the accompanying work by Hossain et al.) provide evidence of the involvement of splicing, translation, and decay in the regulation of early events in mRNP biogenesis; and imply the additional requirement for a balance in splicing isoforms from a single gene.
mRNA 合成与输出的有效偶联对于基因表达至关重要。Sus1/ENY2 是 SAGA 和 TREX-2 复合物的一个组成部分,参与转录和 mRNA 输出。虽然大多数酵母基因缺乏内含子,但我们之前曾报道过酵母 SUS1 带有两个内含子。在这里,我们表明这种特征在进化上是保守的,对 Sus1 功能至关重要。我们确定虽然 SUS1 的剪接效率不高,但它会响应细胞条件,促进或阻断剪接的内含子突变会导致 mRNA 输出和细胞生长缺陷。与此一致,我们发现无内含子的 SUS1 只能部分挽救 sus1Δ 表型。值得注意的是,每个 SUS1 内含子的剪接也受到另一个内含子和 SUS1 外显子序列的影响。此外,通过跟踪 SUS1 RNA 和蛋白质水平,我们确定了无意义介导的衰变 (NMD) 途径和剪接因子 Mud2 都在 SUS1 表达中发挥作用。我们的数据(以及 Hossain 等人的相关工作)提供了证据,证明剪接、翻译和降解参与了 mRNP 生物发生早期事件的调节;并暗示需要平衡单个基因的剪接异构体。