Yeh Tsung-Han, Huang Shu-Yu, Lan Wan-Yu, Liaw Gwo-Jen, Yu Jenn-Yah
Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Taipei, Taiwan.
Brain Research Center, National Yang-Ming University, Taipei, Taiwan.
Dev Dyn. 2015 Jul;244(7):852-65. doi: 10.1002/dvdy.24292. Epub 2015 Jun 22.
Tousled-like kinase (Tlk) is a conserved serine/threonine kinase regulating DNA replication, chromatin assembly, and DNA repair. Previous studies have suggested that Tlk is involved in cell morphogenesis in vitro. In addition, tlk genetically interact with Rho1, which encodes a key regulator of the cytoskeleton. However, whether Tlk plays a physiological role in cell morphogenesis and cytoskeleton rearrangement remains unknown.
In tlk mutant follicle cells, area of the apical domain was reduced. The density of microtubules was increased in tlk mutant cells. The density of actin filaments was increased in the apical region and decreased in the basal region. Because area of the apical domain was reduced, we examined the levels of proteins located in the apical region by using immunofluorescence. The fluorescence intensities of two adherens junction proteins Armadillo (Arm) and DE-cadherin (DE-cad), atypical protein kinase C (aPKC), and Notch, were all increased in tlk mutant cells. The basolateral localized Discs large (Dlg) shifted apically in tlk mutant cells.
Increase of protein densities in the apical region might be resulted from disruption of the cytoskeleton and shrinkage of the apical domain. Together, these data suggest a novel role of Tlk in maintaining cell morphology, possibly through modulating the cytoskeleton.
类蓬松激酶(Tlk)是一种保守的丝氨酸/苏氨酸激酶,可调节DNA复制、染色质组装和DNA修复。先前的研究表明,Tlk在体外参与细胞形态发生。此外,tlk与Rho1发生遗传相互作用,Rho1编码细胞骨架的关键调节因子。然而,Tlk是否在细胞形态发生和细胞骨架重排中发挥生理作用仍不清楚。
在tlk突变的卵泡细胞中,顶端区域面积减小。tlk突变细胞中微管密度增加。肌动蛋白丝密度在顶端区域增加,在基部区域减少。由于顶端区域面积减小,我们通过免疫荧光检测了位于顶端区域的蛋白质水平。两种黏着连接蛋白犰狳蛋白(Arm)和DE-钙黏蛋白(DE-cad)、非典型蛋白激酶C(aPKC)和Notch的荧光强度在tlk突变细胞中均增加。基底外侧定位的盘状大蛋白(Dlg)在tlk突变细胞中向顶端移位。
顶端区域蛋白质密度的增加可能是由于细胞骨架破坏和顶端区域收缩所致。总之,这些数据表明Tlk在维持细胞形态方面具有新作用,可能是通过调节细胞骨架实现的。