Kim Byung-Hoon, Cai Xue, Vaughn Justin N, von Arnim Albrecht G
Department of Biochemistry, Cellular and Molecular Biology, The University of Tennessee, Knoxville, TN 37996-0840, USA.
Genome Biol. 2007;8(4):R60. doi: 10.1186/gb-2007-8-4-r60.
The eukaryotic translation initiation factor 3 (eIF3) has multiple roles during the initiation of translation of cytoplasmic mRNAs. How individual subunits of eIF3 contribute to the translation of specific mRNAs remains poorly understood, however. This is true in particular for those subunits that are not conserved in budding yeast, such as eIF3h.
Working with stable reporter transgenes in Arabidopsis thaliana mutants, it was demonstrated that the h subunit of eIF3 contributes to the efficient translation initiation of mRNAs harboring upstream open reading frames (uORFs) in their 5' leader sequence. uORFs, which can function as devices for translational regulation, are present in over 30% of Arabidopsis mRNAs, and are enriched among mRNAs for transcriptional regulators and protein modifying enzymes. Microarray comparisons of polysome loading in wild-type and eif3h mutant seedlings revealed that eIF3h generally helps to maintain efficient polysome loading of mRNAs harboring multiple uORFs. In addition, however, eIF3h also boosted the polysome loading of mRNAs with long leaders or coding sequences. Moreover, the relative polysome loading of certain functional groups of mRNAs, including ribosomal proteins, was actually increased in the eif3h mutant, suggesting that regulons of translational control can be revealed by mutations in generic translation initiation factors.
The intact eIF3h protein contributes to efficient translation initiation on 5' leader sequences harboring multiple uORFs, although mRNA features independent of uORFs are also implicated.
真核生物翻译起始因子3(eIF3)在细胞质mRNA翻译起始过程中具有多种作用。然而,eIF3的各个亚基如何促进特定mRNA的翻译仍知之甚少。对于那些在芽殖酵母中不保守的亚基,如eIF3h,尤其如此。
通过在拟南芥突变体中使用稳定的报告转基因,证明了eIF3的h亚基有助于5'前导序列中含有上游开放阅读框(uORF)的mRNA的有效翻译起始。uORF可作为翻译调控的元件,存在于超过30%的拟南芥mRNA中,并且在转录调节因子和蛋白质修饰酶的mRNA中富集。野生型和eif3h突变体幼苗中多核糖体负载的微阵列比较显示,eIF3h通常有助于维持含有多个uORF的mRNA的有效多核糖体负载。然而,此外,eIF3h还提高了具有长前导序列或编码序列的mRNA的多核糖体负载。此外,包括核糖体蛋白在内的某些功能组mRNA在eif3h突变体中的相对多核糖体负载实际上增加了,这表明通用翻译起始因子的突变可以揭示翻译控制的调控子。
完整的eIF3h蛋白有助于在含有多个uORF的5'前导序列上进行有效的翻译起始,尽管也涉及独立于uORF的mRNA特征。