Espunya M C, Combettes B, Dot J, Chaubet-Gigot N, Martínez M C
Departamento de Bioquímica y Biología Molecular, Universidad Autónoma de Barcelona, Spain.
Plant J. 1999 Sep;19(6):655-66. doi: 10.1046/j.1365-313x.1999.00563.x.
Protein kinase CK2 is an ubiquitous Ser/Thr kinase essential for cell growth. We have used the highly synchronizable tobacco BY-2 cell line to investigate whether CK2 activity and expression are regulated in a cell cycle phase-dependent manner in higher plants. Specific cDNA probes for tobacco CK2alpha and beta subunits, respectively, and polyclonal antibodies recognising alpha and beta subunits separately, were obtained to determine mRNA and protein levels of both subunits. Our results show that CK2 activity oscillates throughout the cell cycle, peaking at G1/S and M phases, due to a post-translational regulation of the tetrameric enzyme. Additional levels of control of CK2 expression operate in relation to the proliferative state of the cells, including differential accumulation of alpha and beta transcripts and post-transcriptional regulation of protein levels (beta subunit). Moreover, in vivo inhibition of CK2 activity corroborates the requirement of the functional CK2 to progress through the cell division cycle, and suggests that CK2 might play an important role at the G2/M checkpoint.
蛋白激酶CK2是一种对细胞生长至关重要的普遍存在的丝氨酸/苏氨酸激酶。我们利用高度可同步化的烟草BY-2细胞系来研究在高等植物中CK2的活性和表达是否以细胞周期阶段依赖性方式受到调控。分别获得了针对烟草CK2α和β亚基的特异性cDNA探针,以及分别识别α和β亚基的多克隆抗体,以确定两个亚基的mRNA和蛋白质水平。我们的结果表明,由于四聚体酶的翻译后调控,CK2活性在整个细胞周期中振荡,在G1/S期和M期达到峰值。CK2表达的额外调控水平与细胞的增殖状态相关,包括α和β转录本的差异积累以及蛋白质水平(β亚基)的转录后调控。此外,体内对CK2活性的抑制证实了功能性CK2对细胞分裂周期进程的必要性,并表明CK2可能在G2/M检查点发挥重要作用。