School of Biotechnology and Environmental Engineering, Myongji University, Yongin, Kyunggi-do 449-728, South Korea.
Plant J. 2010 Dec;64(6):960-76. doi: 10.1111/j.1365-313X.2010.04393.x. Epub 2010 Nov 17.
Pumilio, an RNA-binding protein that contains tandemly repeated Puf domains, is known to repress translational activity in early embryogenesis and polarized cells of non-plant species. Although Pumilio proteins have been characterized in many eukaryotes, their role in plants is unknown. In the present study, we characterized an Arabidopsis Pumilio-encoding gene, APUM23. APUM23 is constitutively expressed, with higher levels in metabolically active tissues, and its expression is up-regulated in the presence of either glucose or sucrose. The T-DNA insertion mutants apum23-1 and apum23-2 showed slow growth, with serrated and scrunched leaves, an abnormal venation pattern, and distorted organization of the palisade parenchyma cells - a phenotype that is reminiscent of nucleolin and ribosomal protein gene mutants. Intracellular localization studies indicate that APUM23 predominantly localizes to the nucleolus. Based on this localization, rRNA processing was examined. In apum23, 35S pre-rRNA, and unprocessed 18S and 5.8S poly(A) rRNAs, accumulated without affecting the steady-state levels of mature rRNAs, indicating that APUM23 is involved in the processing and/or degradation of 35S pre-rRNA and rRNA maturation by-products. The apum23 mutant showed increased levels of 18S rRNA biogenesis-related U3 and U14 small nucleolar RNAs (snoRNAs) and accumulated RNAs within the nucleolus. Our data suggest that APUM23 plays an important role in plant development via rRNA processing.
Pumilio 是一种 RNA 结合蛋白,含有串联重复的 Puf 结构域,已知在早期胚胎发生和非植物物种的极化细胞中抑制翻译活性。尽管 Pumilio 蛋白已在许多真核生物中得到表征,但它们在植物中的作用尚不清楚。在本研究中,我们对拟南芥 Pumilio 编码基因 APUM23 进行了表征。APUM23 组成型表达,在代谢活跃的组织中表达水平较高,其表达在存在葡萄糖或蔗糖时上调。T-DNA 插入突变体 apum23-1 和 apum23-2 表现出生长缓慢,叶片锯齿状和皱缩,叶脉模式异常,栅栏薄壁组织细胞的组织扭曲 - 这种表型类似于核仁素和核糖体蛋白基因突变体。细胞内定位研究表明,APUM23 主要定位于核仁。基于这种定位,研究了 rRNA 加工。在 apum23 中,35S 前 rRNA 和未加工的 18S 和 5.8S 多 A rRNAs 积累,而不影响成熟 rRNA 的稳态水平,表明 APUM23 参与 35S 前 rRNA 的加工和/或降解以及 rRNA 成熟的副产物。apum23 突变体显示出 18S rRNA 生物发生相关 U3 和 U14 小核仁 RNA(snoRNA)水平升高,并在核仁内积累 RNA。我们的数据表明,APUM23 通过 rRNA 加工在植物发育中发挥重要作用。