Cytoplasmic relocation of Daxx induced by Ro52 and FLASH.
The RING-finger protein Ro52/TRIM21 is known to be an autoantigen and is recognized by anti-Ro/SSA antibodies, which are commonly found in patients with Sjögren's syndrome and systemic lupus erythematosus. We recently showed that Ro52 is an E3 ubiquitin ligase and localizes to cytoplasmic bodies that are highly motile along the microtubule network. To expand our knowledge of Ro52, we searched partners co-operating with Ro52. We performed a yeast two-hybrid screening of a human brain cDNA library with Ro52 as bait. This screening identified several genes encoding Ro52-interacting proteins, including the apoptosis-related proteins, Daxx and FLASH. Further yeast two-hybrid assays revealed that Daxx binds to the B30.2 domain of Ro52 and that FLASH binds to coiled-coil domains of Ro52 through its death-effector domain-recruiting domain. These results suggest that Ro52, Daxx, and FLASH form heteromeric protein complexes. Indeed, this was supported by results of immunoprecipitation experiments in which we found that Daxx is co-immunoprecipitated with Ro52 in the presence of overexpressed FLASH. Importantly, our fluorescence microscopy revealed that, although Daxx is predominantly located in the nucleus, overexpression of both Ro52 and FLASH leads to relocation of Daxx into the cytoplasm. Thus, Ro52 seems to co-operate with FLASH to induce cytoplasmic localization of Daxx in cells.
Tanaka
Makoto
M
Department of Medicine, Center for Molecular Chaperone/Radiobiology and Cancer Virology, MCG Cancer Center, Medical College of Georgia, 1410 Laney Walker Blvd., CN-3141, Augusta, GA 30912, USA.
Kamitani
Tetsu
T
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D051017:Apoptosis Regulatory Proteins / Q000502:physiology*; D001665:Binding Sites; D002135:Calcium-Binding Proteins / Q000502:physiology*; D045744:Cell Line, Tumor; D056970:Co-Repressor Proteins; D003593:Cytoplasm / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D018832:Molecular Chaperones; D009687:Nuclear Proteins / Q000378:metabolism*; D021381:Protein Transport; D012261:Ribonucleoproteins / Q000502:physiology*; D020798:Two-Hybrid System Techniques; D000099207:SS-A Antigen
D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D002135:Calcium-Binding Proteins; D056970:Co-Repressor Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C485604:CASP8AP2 protein, human; C106985:DAXX protein, human