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Hedgehog信号通路在胚胎发育、组织再生中发挥重要的作用,且与癌症发生发展密切相关.其胞内调控组分Suppressor of Fused(SuFu)蛋白通过结合转录因子Gli(s),负调控该信号通路,但其作用的分子机制仍不甚清楚.在本项研究中,以人SuFu作为诱饵蛋白,利用酵母双杂交技术成功地筛选到1个新的相互作用因子—蛋白酶体成熟蛋白(POMP).通过免疫共沉淀、体外GST pull-down和免疫细胞化学实验验证其相互作用.为了探究POMP与SuFu的相互作用对Hedgehog信号通路的影响,构建了POMP的过表达质粒和干扰质粒(miR-RNAi)以及转录因子Gli活性检测系统,即荧光素酶报告基因法,结果显示,过表达SuFu蛋白时POMP正调控Hedgehog信号通路,而下调POMP的表达则抑制Gli的活性.该研究结果揭示了POMP新的生物学功能,为阐明Hedgehog信号通路的具体分子机制提供了新的线索.
The Hedgehog signaling pathway plays an important role in embryonic development and tissue regeneration and is closely related to the development of cancer. Suppressor of Fused (SuFu) protein negatively regulates this signaling pathway by binding transcription factor Gli (s) , But the molecular mechanism of its action remains unclear.In this study, a new interacting factor, proteasome maturation protein (POMP), was successfully screened by human yeast SuFu as a bait protein and yeast two-hybrid technique. The interaction between POMP and SuFu in Hedgehog signaling pathway was investigated by co-immunoprecipitation, GST pull-down and immunocytochemistry in vitro. To investigate the effect of POMP and SuFu on the Hedgehog signaling pathway, we constructed the overexpression plasmid and miR- And luciferase reporter assay, a transcription factor Gli activity assay, showed that POMP positively regulated the Hedgehog signaling pathway when overexpressing SuFu protein while inhibiting the expression of POMP inhibited Gli activity.The results of this study revealed that POMP Biological function, providing new clues for clarifying the specific molecular mechanism of the Hedgehog signaling pathway.