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目的：研究Ppp5c 及TTC16 基因的TPR (tetratricopeptide repeat)结构域短片段和Hsp70 及Hsp90 家族蛋白的相互做用，及其过表达对细胞周期的影响。方法：通过生物信息学的分析及PCR 的方法，克隆Ppp5c 及TTC16 基因的TPR 结构域以及 HSPA1A、HSP90AA1 的全长基因，并连入酵母双杂交载体，通过ClonTech 的酵母双杂交实验体系研究蛋白和蛋白之间的相互作用。把Ppp5c 及TTC16 基因的TPR 结构域克隆入真核表达载体，构架稳定表达Ppp5c 及TTC16 基因的TPR 结构域的MCF-7 细胞系，并通过流式细胞实验观察细胞周期。结果：Ppp5c 及TTC16 基因的TPR 结构域能与HSPA1A 或HSP90AA1 发生相互作用。 Ppp5c 及TTC16 基因的TPR 结构域在MCF-7 中的过表达能严重影响细胞周期，引起细胞凋亡和S 期阻滞。结论：本实验初步揭示了不同蛋白的TPR 结构域在与Hsp70 及Hsp90 蛋白的相互作用性质的异同点以及其过表达对细胞周期的影响，为全面理解 TPR 结构域的功能、Ppp5c 以及TTC16 蛋白在细胞内的功能奠定了前期实验基础。

英文摘要:

Objective: To investigate the interactions between the TPR (tetratricopeptide repeat) domains of Ppp5c/TTC16 and protein of Hsp70/Hsp90, to study the effect of the overexpression of TPR domains of Ppp5c/TTC16 on cell cycle in MCF-7 cell lines. Methods: After being analyzed by online informatics tools, the TPR domains of Ppp5c/TTC16 and full-length of HSPA1A/HSP90AA1 genes were respectively sub-cloned into two separate vectors of the yeast two-hybrid system provided by ClonTech to confirm the protein-protein interactions. The TPR domains of Ppp5c/TTC16 were sub-cloned into eukaryotic expression vectors which were stably transfected into MCF-7 cell-lines, whose cell cycle was then examined by flow cytometry. Results: The TPR domains of Ppp5c/TTC16 interacted with proteins of HSPA1A/HSP90AA1. A large amount of apoptosis was detected when the TPR domains of Ppp5c over-expressed in MCF-7 cell line, and the cell cycle was blocked at the S phase when the TPR domains of TTC16 over-expressed in MCF-7 cell line. Conclusion: This research revealed the discrepancy in the capacity of the combination of dissimilar TPR domains with Hsp70/Hsp90 proteins, as well as the effect of overexpression of TPR domains on the cell-cycle of MCF-7 cell line, which may help draw the panoramic picture of the functions of TPR domains and deep the understanding of intra-cellular functions of Ppp5c and TTC16 proteins.

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