Jeon Young, Park Yong-Joon, Cho Hui Kyung, Jung Hyun Ju, Ahn Tae-Kyu, Kang Hunseung, Pai Hyun-Sook
Department of Systems Biology, Yonsei University, Seoul 120-749, Korea.
Department of Plant Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 500-757, Korea.
J Exp Bot. 2015 Oct;66(20):6297-310. doi: 10.1093/jxb/erv337. Epub 2015 Jul 10.
Nucleostemin is a nucleolar GTP-binding protein that is involved in stem cell proliferation, embryonic development, and ribosome biogenesis in mammals. Plant nucleostemin-like 1 (NSN1) plays a role in embryogenesis, and apical and floral meristem development. In this study, a nucleolar function of NSN1 in the regulation of ribosome biogenesis was identified. Green fluorescent protein (GFP)-fused NSN1 localized to the nucleolus, which was primarily determined by its N-terminal domain. Recombinant NSN1 and its N-terminal domain (NSN1-N) bound to RNA in vitro. Recombinant NSN1 expressed GTPase activity in vitro. NSN1 silencing in Arabidopsis thaliana and Nicotiana benthamiana led to growth retardation and premature senescence. NSN1 interacted with Pescadillo and EBNA1 binding protein 2 (EBP2), which are nucleolar proteins involved in ribosome biogenesis, and with several ribosomal proteins. NSN1, NSN1-N, and EBP2 co-fractionated primarily with the 60S ribosomal large subunit in vivo. Depletion of NSN1 delayed 25S rRNA maturation and biogenesis of the 60S ribosome subunit, and repressed global translation. NSN1-deficient plants exhibited premature leaf senescence, excessive accumulation of reactive oxygen species, and senescence-related gene expression. Taken together, these results suggest that NSN1 plays a crucial role in plant growth and senescence by modulating ribosome biogenesis.
核仁素是一种核仁鸟苷三磷酸结合蛋白,参与哺乳动物的干细胞增殖、胚胎发育和核糖体生物合成。植物核仁素样蛋白1(NSN1)在胚胎发生以及顶端和花分生组织发育中发挥作用。在本研究中,确定了NSN1在核糖体生物合成调控中的核仁功能。绿色荧光蛋白(GFP)融合的NSN1定位于核仁,这主要由其N端结构域决定。重组NSN1及其N端结构域(NSN1-N)在体外与RNA结合。重组NSN1在体外表现出GTPase活性。在拟南芥和本氏烟草中沉默NSN1导致生长迟缓以及早衰。NSN1与参与核糖体生物合成的核仁蛋白Pescadillo和EBNA1结合蛋白2(EBP2)相互作用,还与几种核糖体蛋白相互作用。在体内,NSN1、NSN1-N和EBP2主要与60S核糖体大亚基共分离。NSN1的缺失延迟了25S rRNA的成熟以及60S核糖体亚基的生物合成,并抑制了整体翻译。NSN1缺陷型植物表现出叶片早衰、活性氧的过度积累以及衰老相关基因的表达。综上所述,这些结果表明NSN1通过调节核糖体生物合成在植物生长和衰老中起关键作用。