Division of Evolution & Genomic Sciences, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK.
Laboratorio de Genética y Enfermedades Metabólicas, Instituto de Nutrición y Tecnología de los Alimentos (INTA), Universidad de Chile, Santiago, Chile.
Nat Commun. 2021 Feb 5;12(1):833. doi: 10.1038/s41467-021-21053-2.
The structure of proline prevents it from adopting an optimal position for rapid protein synthesis. Poly-proline-tract (PPT) associated ribosomal stalling is resolved by highly conserved eIF5A, the only protein to contain the amino acid hypusine. We show that de novo heterozygous EIF5A variants cause a disorder characterized by variable combinations of developmental delay, microcephaly, micrognathia and dysmorphism. Yeast growth assays, polysome profiling, total/hypusinated eIF5A levels and PPT-reporters studies reveal that the variants impair eIF5A function, reduce eIF5A-ribosome interactions and impair the synthesis of PPT-containing proteins. Supplementation with 1 mM spermidine partially corrects the yeast growth defects, improves the polysome profiles and restores expression of PPT reporters. In zebrafish, knockdown eif5a partly recapitulates the human phenotype that can be rescued with 1 µM spermidine supplementation. In summary, we uncover the role of eIF5A in human development and disease, demonstrate the mechanistic complexity of EIF5A-related disorder and raise possibilities for its treatment.
脯氨酸的结构使其无法采用最适合快速蛋白质合成的位置。多脯氨酸结构域(PPT)相关的核糖体stalling 通过高度保守的 eIF5A 解决,eIF5A 是唯一含有氨基酸 hypusine 的蛋白质。我们表明,新的杂合性 EIF5A 变体导致一种疾病,其特征是发育迟缓、小头症、小颌畸形和发育异常的组合不同。酵母生长测定、多核糖体分析、总/hypusinated eIF5A 水平和 PPT 报告基因研究表明,这些变体损害了 eIF5A 的功能,减少了 eIF5A-核糖体的相互作用,并损害了含有 PPT 的蛋白质的合成。补充 1mM 亚精胺部分纠正了酵母生长缺陷,改善了多核糖体图谱,并恢复了 PPT 报告基因的表达。在斑马鱼中,eif5a 的敲低部分重现了人类表型,用 1μM 亚精胺补充可以挽救该表型。总之,我们揭示了 eIF5A 在人类发育和疾病中的作用,证明了 EIF5A 相关疾病的机制复杂性,并为其治疗提供了可能性。