Gao Chun, Negash Sewite, Guo Hong Tao, Ledee Dolena, Wang Hwai-Shi, Zelenka Peggy
National Eye Institute, NIH, Bethesda, MD. Yang Ming University, Taipei, Taiwan.
Mol Cancer Res. 2002 Nov;1(1):12-24.
CDK5 and its activator, p35, are expressed in mouse corneal epithelium and can be coimmunoprecipited from corneal epithelial cell lysates. Immunostaining shows CDK5 and p35 in all layers of the corneal epithelium, especially along the basal side of the basal cells. Stable transfection of corneal epithelial cells with CDK5, which increases CDK5 kinase activity by approximately 33%, also increases the number of cells adhering to fibronectin and the strength of adhesion. CDK5 kinase activity seems to be required for this effect, because the kinase inactive mutation, CDK5-T33, either reduces adhesion or has no significant effect, depending on the level of expression. Using an in vitro scrape wound in confluent cultures of stably transfected cells to examine the effect of CDK5 on cell migration, we show that reoccupation of the wound area is significantly decreased by CDK5 and increased by CDK5-T33. These findings indicate that CDK5 may be an important regulator of adhesion and migration of corneal epithelial cells.
细胞周期蛋白依赖性激酶5(CDK5)及其激活剂p35在小鼠角膜上皮中表达,并且可以从小鼠角膜上皮细胞裂解物中共免疫沉淀出来。免疫染色显示CDK5和p35存在于角膜上皮的所有层中,尤其是在基底细胞的基底侧。用CDK5稳定转染角膜上皮细胞,可使CDK5激酶活性增加约33%,同时也增加了细胞与纤连蛋白的黏附数量和黏附强度。这种作用似乎需要CDK5激酶活性,因为激酶失活突变体CDK5-T33根据表达水平不同,要么降低黏附力,要么无显著影响。利用稳定转染细胞的汇合培养物中的体外划痕伤口来检测CDK5对细胞迁移的影响,我们发现,CDK5会显著减少伤口区域的重新占据,而CDK5-T33则会增加伤口区域的重新占据。这些发现表明,CDK5可能是角膜上皮细胞黏附和迁移的重要调节因子。