摘要
The Hippo pathway is evolutionarily conserved from Drosophila to mammals. FRMD6 is one crucial up- stream component of the Hippo pathway while its function and regulatory mechanism are largely elusive. We decided to purify the protein complex of FRMD6 to further explore its regulatory mechanism. We established the MCF7 breast cancer cells that stably expressed FRMD6 by retroviruses infection. And we purified the FRMD6-interacting protein complex for further analysis by high performance liquid chromatography-mass spectrometry/mass spectrometry（HPLC- MS/MS）. Interestingly, we observed that the major binding partner of FRMD6 is 14-3-3 family of proteins. The interac- tion between FRMD6 and 14-3-3 proteins was detected by co-immunoprecipitation（CO-IP）. The disruption of their interaction resulted in the nuclear localization of FRMD6. Importantly, the T28A mutant of FRMD6 showed stronger tumor suppressor function than wild type（WT） FRMD6. Our results indicate that 14-3-3 proteins tightly regulate the subcellular localization of FRMD6 so as to endow FRMD6 with the tumor suppressor function on breast cancer.
The Hippo pathway is evolutionarily conserved from Drosophila to mammals. FRMD6 is one crucial up- stream component of the Hippo pathway while its function and regulatory mechanism are largely elusive. We decided to purify the protein complex of FRMD6 to further explore its regulatory mechanism. We established the MCF7 breast cancer cells that stably expressed FRMD6 by retroviruses infection. And we purified the FRMD6-interacting protein complex for further analysis by high performance liquid chromatography-mass spectrometry/mass spectrometry（HPLC- MS/MS）. Interestingly, we observed that the major binding partner of FRMD6 is 14-3-3 family of proteins. The interac- tion between FRMD6 and 14-3-3 proteins was detected by co-immunoprecipitation（CO-IP）. The disruption of their interaction resulted in the nuclear localization of FRMD6. Importantly, the T28A mutant of FRMD6 showed stronger tumor suppressor function than wild type（WT） FRMD6. Our results indicate that 14-3-3 proteins tightly regulate the subcellular localization of FRMD6 so as to endow FRMD6 with the tumor suppressor function on breast cancer.
基金
Supported by the National Natural Science Foundation of China（No.30973274） and the Doctoral Fund of Youth Scholars of Ministry of Education of China（No.20090061120073）
