Institute of Natural Sciences, Senshu University, Higashimita 2-1-1, Tama, Kawasaki, Kanagawa, 214-8580, Japan.
Department of Life Science, College of Science, Rikkyo University, Toshima, Tokyo, 171-8501, Japan.
J Plant Res. 2024 Nov;137(6):1091-1104. doi: 10.1007/s10265-024-01578-5. Epub 2024 Sep 5.
Nucleotides are the building blocks of living organisms and their biosynthesis must be tightly regulated. Inosine monophosphate dehydrogenase (IMPDH) is a rate-limiting enzyme in GTP synthesis that is essential for biological activities, such as RNA synthesis. In animals, the suppression of IMPDH function causes ribosomal stress (also known as nucleolar stress), a disorder in ribosome biogenesis that results in cell proliferation defects and apoptosis. Despite its importance, plant IMPDH has not been analyzed in detail. Therefore, we analyzed the phenotypes of mutants of the two IMPDH genes in Arabidopsis thaliana and investigated their relationship with ribosomal stress. Double mutants of IMPDH1 and IMPDH2 were lethal, and only the impdh2 mutants showed growth defects and transient chlorophyll deficiency. These results suggested that IMPDH1 and IMPDH2 are redundant and essential, whereas IMPDH2 has a crucial role. In addition, the impdh2 mutants showed a reduction in nucleolus size and resistance to several translation inhibitors, which is a known response to ribosomal stress. Furthermore, the IMPDH1/impdh1 impdh2 mutants showed more severe growth defects and phenotypes such as reduced plastid rRNA levels and abnormal processing patterns than the impdh2 mutants. Finally, multiple mutations of impdh with as2, which has abnormal leaf polarity, caused the development of needle-like leaves because of the enhancement of the as2 phenotype, which is a typical effect observed in mutants of genes involved in ribosome biogenesis. These results indicated that IMPDH is closely related to ribosome biogenesis, and that mutations in the genes lead to not only known responses to ribosomal stress, but also plant-specific responses.
核苷酸是生物的构建模块,其生物合成必须受到严格的调控。肌苷单磷酸脱氢酶(IMPDH)是 GTP 合成的限速酶,对于 RNA 合成等生物活性至关重要。在动物中,抑制 IMPDH 的功能会导致核糖体应激(也称为核仁应激),这是一种核糖体生物发生失调的疾病,导致细胞增殖缺陷和细胞凋亡。尽管其重要性,但植物 IMPDH 尚未得到详细分析。因此,我们分析了拟南芥中两种 IMPDH 基因的突变体的表型,并研究了它们与核糖体应激的关系。IMPDH1 和 IMPDH2 的双突变体是致死的,只有 impdh2 突变体表现出生长缺陷和短暂的叶绿素缺乏。这些结果表明 IMPDH1 和 IMPDH2 是冗余的和必需的,而 IMPDH2 则起着关键作用。此外,impdh2 突变体的核仁大小减小,对几种翻译抑制剂的抗性增强,这是已知的核糖体应激反应。此外,IMPDH1/impdh1 impdh2 双突变体比 impdh2 突变体表现出更严重的生长缺陷和表型,如质体 rRNA 水平降低和异常加工模式。最后,与 as2(具有异常叶极性)的 impdh 多次突变导致针状叶片的发育,因为 as2 表型的增强,这是核糖体生物发生基因突变体中观察到的典型效应。这些结果表明 IMPDH 与核糖体生物发生密切相关,基因突变不仅导致已知的核糖体应激反应,还导致植物特异性反应。