Characterization Of The Biochemical Properties Of LMX1B In Human Keratinocytes

• Main Authors: Hsu,Chiachen, 許家禎
• Other Authors: Huang,Shihming
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/71361312408045212595

碩士 === 國防醫學院 === 生物化學研究所 === 100 === The nail-patella syndrome (NPS) is a rare autosomal-dominant disease, and it causes nails and bones dysplasia. Recent evidence shows that NPS is the result of mutations in the LMX1B transcription factor, and different LMX1B mutations will cause the diversity of NPS clinical results. Therefore, the role of LMX1B in NPS still remains unclear. Interestingly, LMX1B can induce the transcription of IL-6, a NF-kB target gene, in HeLa cells. Moreover, 95% of patients have dysplastic nails and K17 is found highly expressed in normal nail matrix, in which the destruction may induce nail dysplastia. Therefore, in this thesis we will focus on the effect of LMX1B variation on the biochemical properties, and the possible role in the gene expression of IL-6 and K17 in human keratinocytes. By using the fluorescence microscopy, we found a dominant NLS in amino acid sequence 198-202 of LMX1B. Both C-terminal truncated LMX1B R198X and LMX1B R200Q mutants leak out from the nucleus. By using luciferase reporter assay and western blot, we found that the integrity of both homeodomain and LIMB domain are important for the regulation of IL-6 promoter activity and K17 expression. Clinical results show that the location of LMX1B R198X C-terminal truncated and LMX1B R200Q mutation are different from wild-type and their influence to IL-6 and K17may not be the same. However, the influence of the mutation on wild-type LMX1B and whether NPS is affected by K17 negative regulation await further investigations.