Kataya Amr R A, Creighton Maria T, Napitupulu Toga P, Sætre Christine, Heidari Behzad, Ruoff Peter, Lillo Cathrine
Centre for Organelle Research, Faculty of Science and Technology, University of Stavanger, Stavanger, Norway.
PLoS One. 2017 Jul 5;12(7):e0180478. doi: 10.1371/journal.pone.0180478. eCollection 2017.
Eukaryotic protein phosphatase 4 (PP4) is a PP2A-type protein phosphatase that is part of a conserved complex with regulatory factors PSY2 and PP4R2. Various lines of Arabidopsis thaliana with mutated PP4 subunit genes were constructed to study the so far completely unknown functions of PP4 in plants. Mutants with knocked out putative functional homolog of the PSY2 LIKE (PSY2L) gene were dwarf and bushy, while plants with knocked out PP4R2 LIKE (PP4R2L) looked very similar to WT. The psy2l seedlings had short roots with disorganized morphology and impaired meristem. Seedling growth was sensitive to the genotoxin cisplatin. Global transcript analysis (RNA-seq) of seedlings and rosette leaves revealed several groups of genes, shared between both types of tissues, strongly influenced by knocked out PSY2L. Receptor kinases, CRINKLY3 and WAG1, important for growth and development, were down-regulated 3-7 times. EUKARYOTIC ELONGATION FACTOR5A1 was down-regulated 4-6 fold. Analysis of hormone sensitive genes indicated that abscisic acid levels were high, while auxin, cytokinin and gibberellic acid levels were low in psy2l. Expression of specific transcription factors involved in regulation of anthocyanin synthesis were strongly elevated, e.g. the master regulator PAP1, and intriguingly TT8, which is otherwise mainly expressed in seeds. The psy2l mutants accumulated anthocyanins under conditions where WT did not, pointing to PSY2L as a possible upstream negative regulator of PAP1 and TT8. Expression of the sugar-phosphate transporter GPT2, important for cellular sugar and phosphate homeostasis, was enhanced 7-8 times. Several DNA damage response genes, including the cell cycle inhibitor gene WEE1, were up-regulated in psy2l. The activation of DNA repair signaling genes, in combination with phenotypic traits showing aberrant root meristem and sensitivity to the genotoxic cisplatin, substantiate the involvement of Arabidopsis PSY2L in maintenance of genome integrity.
真核生物蛋白磷酸酶4(PP4)是一种PP2A类型的蛋白磷酸酶,它是与调节因子PSY2和PP4R2组成的保守复合物的一部分。构建了各种拟南芥PP4亚基基因突变株,以研究PP4在植物中至今完全未知的功能。敲除PSY2类(PSY2L)基因假定功能同源物的突变体矮小且丛生,而敲除PP4R2类(PP4R2L)的植株看起来与野生型非常相似。psy2l幼苗的根短,形态紊乱,分生组织受损。幼苗生长对基因毒素顺铂敏感。对幼苗和莲座叶进行全局转录分析(RNA测序)发现,两类组织中都有几组基因受到PSY2L敲除的强烈影响。对生长和发育很重要的受体激酶CRINKLY3和WAG1下调了3至7倍。真核生物延伸因子5A1下调了4至6倍。对激素敏感基因的分析表明,psy2l中的脱落酸水平较高,而生长素、细胞分裂素和赤霉素水平较低。参与花青素合成调控的特定转录因子的表达大幅升高,例如主要调节因子PAP1,有趣的是还有TT8,否则它主要在种子中表达。psy2l突变体在野生型未积累花青素的条件下积累了花青素,这表明PSY2L可能是PAP1和TT8的上游负调节因子。对细胞糖和磷酸盐稳态很重要的糖磷酸转运蛋白GPT2的表达增强了7至8倍。包括细胞周期抑制基因WEE1在内的几个DNA损伤反应基因在psy2l中上调。DNA修复信号基因的激活,以及显示异常根分生组织和对基因毒性顺铂敏感的表型特征,证实了拟南芥PSY2L参与基因组完整性的维持。