[Analysis of nuclear localization and signal function of XXXX protein predisposing to Warrdenburg syndrome].

@entity10244 @entity2631 @entity3297 @entity22835 @entity22836
OBJECTIVE: To study the role of dysfunction of @entity2631 ( @entity2631 ) of @entity22836 protein in the pathogenesis of @entity10244 . METHODS: Eukaryotic expression plasmid pCMV- @entity22836 -Flag was used as a template to generate mutant plasmid pCMV- @entity22836   @entity2631 -Flag by molecular cloning technique in order to design the mutagenic primers. The UACC903 cells were transfected transiently with @entity22836 and @entity22836   @entity2631 plasmids, and the luciferase activity assays were performed to determine their impact on the transcriptional activities of target gene @entity22835 ( @entity22835 ). The oligonucleotide 5'-GAACGAAGAAGAAGATTT-3' was subcloned into pEGFP-N1 to generate recombinant eukaryotic expression plasmid pEGFP-N1- @entity22836 - @entity2631 . The NIH3T3 cells were transfected separately with @entity22836 , @entity22836   @entity2631 , pEGFP-N1 and pEGFP-N1- @entity2631 plasmids, and their subcellular distribution was observed by immunoflorescence assays. RESULTS: Expression plasmids for the mutant @entity22836   @entity2631 with loss of core @entity2631 sequence and pEGFP-N1- @entity2631 coupled with @entity22836   @entity2631 were successfully generated. Compared with the wild-type @entity22836 , @entity22836   @entity2631 was not able to transactivate the transcriptional activities of promoter @entity22835 and did not affect the normal function of @entity22836 . @entity22836   @entity2631 was only localized in the cytoplasm and pEGFP-N1 was found in both the cytoplasm and nucleus, whereas pEGFP-N1- @entity2631 was mainly located in the nucleus. CONCLUSION: This study has confirmed the localization function of @entity2631 sequence 213ERRRRF218 within the @entity22836 protein. Mutant @entity22836 with loss of @entity2631 has failed to transactivate the transcriptional activities of target gene @entity22835 , which can result in melanocyte defects and cause @entity3297 .
