College of Life Sciences, Hunan University, Changsha, China.
Plant Biol (Stuttg). 2020 Jan;22 Suppl 1:143-152. doi: 10.1111/plb.12954. Epub 2019 Jan 28.
Plastid casein kinase 2 (CK2), which is a major Ser/Thr-specific enzyme in higher organisms, plays an essential role in plant development and diverse abiotic stresses. CKB1 is a regulatory subunit beta of CK2. To expand our understand of functions of the CKB1 gene in Arabidopsis thaliana, protein changes among wild-type (WT) and CKB1 gain- and loss-of-function mutants were compared. Proteins extracted from the CKB1 knockout mutant and overexpressing mutant were compared with Col-0 plants using 2D-PAGE. Proteins regulated by CKB1 were identified with matrix-assisted laser desorption ionisation time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF), and its transcript was verified by qRT-PCR. Bioinformatics analysis, including gene ontology and protein-protein interaction analysis, were employed. The results of mass spectra and bioinformatics analysis suggest that CKB1 may have functions in regulation of the ribosomal protein L10 (RPL10) family and is involved in ultraviolet-B (UV-B) response. Furthermore, qRT-PCR verification showed CKB1 expression was up-regulated by UV-B stress. The expression levels of five genes in the RPL10 family were reduced in the ckb1 T-DNA insertion mutants, whereas they increased in the CKB1 overexpressing mutants under both normal conditions and UV-B treatment. In conclusion, CKB1 has important functions in UV-B radiation stress. Our study implies that CKB1 positively regulates UV-B radiation stress signalling, possibly through modulating expression of the RPL10 family.
质体酪蛋白激酶 2(CK2)是高等生物中主要的丝氨酸/苏氨酸特异性酶,在植物发育和多种非生物胁迫中起着重要作用。CKB1 是 CK2 的调节亚基β。为了扩展我们对拟南芥 CKB1 基因功能的理解,比较了野生型(WT)和 CKB1 功能获得和功能丧失突变体之间的蛋白质变化。使用 2D-PAGE 比较从 CKB1 敲除突变体和过表达突变体中提取的蛋白质与 Col-0 植物的蛋白质。使用基质辅助激光解吸电离飞行时间/飞行时间质谱(MALDI-TOF/TOF)鉴定受 CKB1 调节的蛋白质,并通过 qRT-PCR 验证其转录物。进行了生物信息学分析,包括基因本体和蛋白质-蛋白质相互作用分析。质谱和生物信息学分析的结果表明,CKB1 可能在核糖体蛋白 L10(RPL10)家族的调节中具有功能,并参与紫外线-B(UV-B)反应。此外,qRT-PCR 验证表明,UV-B 胁迫下 CKB1 的表达上调。在 ckb1 T-DNA 插入突变体中,RPL10 家族的五个基因的表达水平降低,而在 CKB1 过表达突变体中,无论是在正常条件下还是在 UV-B 处理下,它们的表达水平都增加。总之,CKB1 在 UV-B 辐射胁迫中具有重要功能。我们的研究表明,CKB1 通过调节 RPL10 家族的表达来正向调节 UV-B 辐射胁迫信号。