人CAP1蛋白与PIF1解螺旋酶及其剪接体蛋白的相互作用

Interaction among human CAP1,PIF1 helicase and its splice variants proteins

目的在细胞水平及体外验证腺苷酸环化酶相关蛋白1(CAP1)蛋白与PIF1解螺旋酶及其剪接体是否存在相互作用。方法采用激光共聚焦免疫荧光实验,通过细胞内蛋白共定位确定蛋白质间的相互作用关系;GST Pulldown实验在体外验证CAP1与PIF1及其剪接体是否存在直接的相互作用。结果激光共聚焦免疫荧光实验显示,CAP1与PIF1全长蛋白及其剪接体BC018978没有共定位,但与剪接体AF108138C存在共定位。GST Pull-down未检测到CAP1与PIF1全长及其剪接体的相互作用。结论 CAP1与PIF1剪接体蛋白可能存在非直接相互作用。

Objective To determine whether human CAP1 interacts with PIF1 helicase and its splice variants proteins in vivo and in vitro. Methods The co-localization of protein was directly observed to determine the interaction between the proteins by confocal immunofluorescence assay. The GST Pull-down experiment was used to test the existence of a direct interaction between CAP1 protein and PIF1 or its splice variants proteins. Results By using confocal immunofluorescence assay, CAP1 protein and PIF1 protein or its splice variants BC018978 had no colocalization, but CAP1 and splice variants AF108138C were co-located in the cytoplasm. Experimental results of the GST Pull-down showed that there was not direct interaction between CAP1 and PIF1 or its splice variants proteins. Conclusion These results show that there may be indirect interaction between CAP1 and PIF1 splice variants proteins.