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Exo70是胞外分泌复合体Exocyst的关键亚基,可通过参与Exocyst复合体或自身聚合促进细胞迁移。我们在研究中发现,Exo70自身形成二聚体的能力显著强于与Exocyst其它7个亚基的结合,提示Exo70二聚化的重要作用。进一步构建Exo70缺失突变体,并通过一系列免疫共沉淀实验确定了Exo70形成二聚体的关键位置是位于氨基酸31~35与505~509的两段同样的序列SLEKS。进而利用这两个序列的缺失突变体,通过transwell迁移实验,证实这两段SLEKS序列在Exo70二聚化及乳腺癌细胞迁移过程中的关键作用。以上结果为Exo70独立于Exocyst复合体功能之外的新功能研究提供了依据,也为乳腺癌及其转移的治疗提供潜在的靶点。
Exo70 is a key subunit of the extracellular secretion complex Exocyst, which promotes cell migration by participating in the Exocyst complex or by self-polymerization. In our study, we found that Exo70 has a significantly stronger ability to dimerize itself than with the other seven subunits of Exocyst, suggesting an important role for Exo70 dimerization. Furthermore, a series of co-immunoprecipitation experiments were carried out to determine whether the Exo70-forming dimer is located at the same two sequences of SLEKS between amino acids 31-35 and 505-509. Furthermore, the deletion mutants of these two sequences were used to confirm the key role of these two SLEKS sequences in the dimerization of Exo70 and the migration of breast cancer cells by transwell migration assay. These results provide a basis for the study of new functions of Exo70 in addition to the functions of Exocyst complex, and provide a potential target for the treatment of breast cancer and its metastasis.