Microphthalmia-associated transcription factor interacts with PU.1 and c-Fos: determination of their subcellular localization.

Marked osteopetrosis is observed only in mi/mi mutant mice although normal osteoclastgenesis is observed in other mutant mice including null mutants at the mi locus. Mutant microphthalmia-associated transcription factor (mi-MITF) has defective nuclear localization potential. We found normal MITF (+-MITF)-c-Fos and mi-MITF-c-Fos complexes in the cytoplasm by immunoblotting, and showed that PU.1 bound with both +-MITF and mi-MITF using an electrophoretic mobility shift assay. Furthermore, the nuclear localization of PU.1 and c-Fos was inhibited by over-expressed mi-MITF in WEHI-3 cells. These results indicate that mi-MITF expressing in osteoclasts specifically binds to c-Fos and PU.1 which are essential transcription factors of osteoclastgenesis and that mi-MITF blocks the nuclear localization of these other transcription factors, which may result in osteopetrosis in mi/mi mutant mice.