Departamento de Bioquímica, Facultad de Química, Universidad Nacional Autónoma de México, Distrito Federal, México.
PLoS One. 2012;7(2):e31606. doi: 10.1371/journal.pone.0031606. Epub 2012 Feb 20.
One of the most regulated steps of translation initiation is the recruitment of mRNA by the translation machinery. In eukaryotes, this step is mediated by the 5'end cap-binding factor eIF4E bound to the bridge protein eIF4G and forming the eIF4F complex. In plants, different isoforms of eIF4E and eIF4G form the antigenically distinct eIF4F and eIF(iso)4F complexes proposed to mediate selective translation. Using a microarray analysis of polyribosome- and non-polyribosome-purified mRNAs from 15 day-old Arabidopsis thaliana wild type [WT] and eIF(iso)4E knockout mutant [(iso)4E-1] seedlings we found 79 transcripts shifted from polyribosomes toward non-polyribosomes, and 47 mRNAs with the opposite behavior in the knockout mutant. The translationally decreased mRNAs were overrepresented in root-preferentially expressed genes and proteins from the endomembrane system, including several transporters such as the phosphate transporter PHOSPHATE1 (PHO1), Sucrose transporter 3 (SUC3), ABC transporter-like with ATPase activity (MRP11) and five electron transporters, as well as signal transduction-, protein modification- and transcription-related proteins. Under normal growth conditions, eIF(iso)4E expression under the constitutive promoter 35 S enhanced the polyribosomal recruitment of PHO1 supporting its translational preference for eIF(iso)4E. Furthermore, under phosphate deficiency, the PHO1 protein increased in the eIF(iso)4E overexpressing plants and decreased in the knockout mutant as compared to wild type. In addition, the knockout mutant had larger root, whereas the 35 S directed expression of eIF(iso)4E caused shorter root under normal growth conditions, but not under phosphate deficiency. These results indicate that selective translation mediated by eIF(iso)4E is relevant for Arabidopsis root development under normal growth conditions.
翻译起始最受调控的步骤之一是翻译机制对 mRNA 的募集。在真核生物中,该步骤由与桥接蛋白 eIF4G 结合的 5'端帽结合因子 eIF4E 介导,并形成 eIF4F 复合物。在植物中,eIF4E 和 eIF4G 的不同同工型形成抗原不同的 eIF4F 和 eIF(iso)4F 复合物,据推测这些复合物介导选择性翻译。我们使用来自 15 天大的拟南芥野生型 [WT] 和 eIF(iso)4E 敲除突变体 [(iso)4E-1] 幼苗的多核糖体和非多核糖体纯化 mRNA 的微阵列分析发现,79 个转录物从多核糖体转移到非多核糖体,并且在敲除突变体中 47 个 mRNA 表现出相反的行为。翻译减少的 mRNA 在根偏好表达的基因和内质网系统中的蛋白质中过度表达,包括几个转运蛋白,如磷酸盐转运蛋白 PHOSPHATE1 (PHO1)、蔗糖转运蛋白 3 (SUC3)、具有 ATP 酶活性的 ABC 转运蛋白样物 (MRP11) 和五个电子转运蛋白,以及信号转导、蛋白质修饰和转录相关蛋白。在正常生长条件下,组成型启动子 35S 下的 eIF(iso)4E 表达增强了 PHO1 的多核糖体募集,支持其对 eIF(iso)4E 的翻译偏好。此外,在磷酸盐缺乏时,与野生型相比,eIF(iso)4E 过表达植物中的 PHO1 蛋白增加,而敲除突变体中的 PHO1 蛋白减少。此外,与野生型相比,敲除突变体的根更大,而在正常生长条件下,35S 指导的 eIF(iso)4E 表达导致根更短,但在磷酸盐缺乏时则不然。这些结果表明,eIF(iso)4E 介导的选择性翻译与拟南芥根在正常生长条件下的发育有关。