Identification of interacting proteins of retinoid-related orphan nuclear receptor gamma in HepG2 cells.
The retinoid-related orphan nuclear receptor gamma (ROR γ) plays critical roles in regulation of development, immunity and metabolism. As transcription factor usually forms a protein complex to function, thus capturing and dissecting of the ROR γ protein complex will be helpful for exploring the mechanisms underlying those functions. After construction of the recombinant tandem affinity purification (TAP) plasmid, pMSCVpuro ROR γ-CTAP(SG), the nuclear localization of ROR γ-CTAP(SG) fusion protein was verified. Following isolation of ROR γ protein complex by TAP strategy, seven candidate interacting proteins were identified. Finally, the heat shock protein 90 (HSP90) and receptor-interacting protein 140 (RIP140) were confirmed to interplay with ROR γ by co-immunoprecipitation. Interference of HSP90 or/and RIP140 genes resulted in dramatically decreased expression of CYP2C8 gene, the ROR γ target gene. Data from this study demonstrate that HSP90 and RIP140 proteins interact with ROR γ protein in a complex format and function as co-activators in the ROR γ-mediated regulatory processes of HepG2 cells.
Institute of Immunology, PLA, Third Military Medical University, Chongqing 400038, China.
Ze-Min
ZM
Huang
Jun
J
Wu
Zheng-Cai
ZC
Jia
Yi
Y
Tian
Jun
J
Tang
Yan
Y
Tang
Ying
Y
Wang
Yu-Zhang
YZ
Wu
Bing
B
Ni
D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D065727:Cytochrome P-450 CYP2C8; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D056971:Nuclear Receptor Co-Repressor 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D018841:HSP90 Heat-Shock Proteins; D056971:Nuclear Receptor Co-Repressor 1; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D010455:Peptides; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C009862:connective tissue-activating peptide; D001189:Aryl Hydrocarbon Hydroxylases; C450259:CYP2C8 protein, human; D065727:Cytochrome P-450 CYP2C8