Departamento de Bioquímica y Biología Molecular, Universitat de Valencia, Burjassot, Spain.
BMC Genet. 2012 Sep 10;13:80. doi: 10.1186/1471-2156-13-80.
The various steps of mRNP biogenesis (transcription, processing and export) are interconnected. It has been shown that the transcription machinery plays a pivotal role in mRNP assembly, since several mRNA export factors are recruited during transcription and physically interact with components of the transcription machinery. Although the shuttling DEAD-box protein Dbp5p is concentrated on the cytoplasmic fibrils of the NPC, previous studies demonstrated that it interacts physically and genetically with factors involved in transcription initiation.
We investigated the effect of mutations affecting various components of the transcription initiation apparatus on the phenotypes of mRNA export mutant strains. Our results show that growth and mRNA export defects of dbp5 and mex67 mutant strains can be suppressed by mutation of specific transcription initiation components, but suppression was not observed for mutants acting in the very first steps of the pre-initiation complex (PIC) formation.
Our results indicate that mere reduction in the amount of mRNP produced is not sufficient to suppress the defects caused by a defective mRNA export factor. Suppression occurs only with mutants affecting events within a narrow window of the mRNP biogenesis process. We propose that reducing the speed with which transcription converts from initiation and promoter clearance to elongation may have a positive effect on mRNP formation by permitting more effective recruitment of partially-functional mRNP proteins to the nascent mRNP.
mRNA 核蛋白颗粒(mRNP)的生物发生(转录、加工和输出)的各个步骤相互关联。已经表明,转录机制在 mRNP 组装中起着关键作用,因为在转录过程中会募集几种 mRNA 输出因子,并且与转录机制的成分物理相互作用。尽管穿梭 DEAD -box 蛋白 Dbp5p 集中在 NPC 的细胞质纤维上,但先前的研究表明,它与参与转录起始的因子在物理和遗传上相互作用。
我们研究了影响转录起始装置的各种成分的突变对 mRNA 输出突变株表型的影响。我们的结果表明,dbp5 和 mex67 突变株的生长和 mRNA 输出缺陷可以通过突变特定的转录起始成分来抑制,但在起始前复合物(PIC)形成的最初步骤中起作用的突变体没有观察到抑制。
我们的结果表明,仅仅减少产生的 mRNP 的数量不足以抑制由有缺陷的 mRNA 输出因子引起的缺陷。仅在影响 mRNP 生物发生过程的狭窄窗口内的事件的突变体中才会发生抑制。我们提出,通过允许部分功能的 mRNP 蛋白更有效地募集到新生的 mRNP,降低转录从起始和启动子清除到延伸的速度可能对 mRNP 形成有积极影响。