摘要
目的探讨ECRG2基因调控中心体复制的分子机制。方法采用质谱法筛选ECRG2基因作用伙伴;采用免疫共沉淀法鉴定ECRG2与p53发生相互作用的位点;采用Western印迹法检测p53转录活性;采用免疫荧光技术观察ECRG2基因对p53中心体定位能力的影响。结果结果显示p53是ECRG2基因的一个新的作用伙伴。ECRG2基因通过其N端20个肽与p53发生相互作用,通过p53参与中心体复制。ECRG2基因缺失不但促进p53蛋白泛素化降解程度,降低p21蛋白活性,提高cyclinE/CDK2活性,从而启动了中心体过度复制,而且使p53蛋白无法定位于中心体,丧失了对中心体再复制的抑制作用。结论ECRG2基因通过p53参与中心体复制,一方面,ECRG2通过p53/p21/cyclin E/CDK2途径调控中心体复制;另一方面,ECRG2也可影响p53中心体定位能力,参与p53对中心体再复制的抑制作用。
Objective To explore the molecular mechanism of ECRG2 participating in regulation of centrosome amplification. Methods The ECRG2-associated proteins were determined using MS analysis, the binding domain between ECRG2 and p53 was identified using inmaunoprecipitation, p53 transactivity was detected by Western blotting, and the localization ability of p53 to centrosomes was measured by inmaunofluesences. Results The results showed that ECRG2 participated in centrosome amplification in a p53-dependent manner. Depletion of ECRG2 not only initiated eentrosome amplification by destabilizing p53, down-regulating p21, and increasing the eyelin E/CDK2 aetivity,but also abolished the localization ability of p53 to eentrosomes. Conclusion The results indicate that ECRG2 partieipates eentrosome duplication through p53 and is important for ensuring eentrosome duplication, and its depletion may contribute to ehromosome instability and aneuploidy in human cancers.
出处
《山西医科大学学报》
CAS
2008年第8期677-681,共5页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(30500588)
山西省自然科学基金资助项目(20051089)
