Identification of interacting proteins of retinoid-related orphan nuclear receptor gamma in HepG2 cells

Identification of interacting proteins of retinoid-related orphan nuclear receptor gamma in HepG2 cells

Blog Article

The retinoid-related orphan nuclear receptor gamma (RORγ)plays critical roles in regulation of development, immunity andmetabolism.As transcription factor usually forms a proteincomplex to function, thus capturing and dissecting of theRORγ protein complex will be helpful for exploring Hay bags themechanisms underlying those functions.After construction ofthe recombinant tandem affinity purification (TAP) plasmid,pMSCVpuro RORγ-CTAP(SG), the nuclear localization ofRORγ-CTAP(SG) fusion protein was verified.

Followingisolation of RORγ protein complex by TAP strategy, sevencandidate interacting proteins were identified.Finally, the heatshock protein Damiana 90 (HSP90) and receptor-interacting protein 140(RIP140) were confirmed to interplay with RORγ byco-immunoprecipitation.Interference of HSP90 or/and RIP140genes resulted in dramatically decreased expression ofCYP2C8 gene, the RORγ target gene.

Data from this studydemonstrate that HSP90 and RIP140 proteins interact withRORγ protein in a complex format and function asco-activators in the RORγ-mediated regulatory processes ofHepG2 cells.