Cloning and characterization of SDPR and PTRF

• Main Authors: Yan-Da Lai, 賴彥達
• Other Authors: Jim C. Fong
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/77830139233429186836

碩士 === 國立陽明大學 === 生化暨分子生物研究所 === 94 === Previously, our lab used 3T3-L1 mouse adipocyte as an experimental model to search for genes that are influenced in insulin resistance. Using suppression subtractive hybridization and RT-PCR, we found serum deprivation response gene (SDPR) was down-regulated by treatment with insulin, TNF-α and isoproterenol. SDPR was a phosphatidylserine binding protein and was up-regulated in serum starvation. GST-pull down analysis indicated that polymerase I and transcript release factor (PTRF) may associate with SDPR. PTRF was shown to play an important role in rRNA transcription termination. In this thesis, we focused on investigating the interaction between SDPR and PTRF in cancer cell line. We found SDPR interacted with PTRF in vivo by using co-immumoprecipitation and mammalian two hybrid assay. In addition, they were co-localizated showed by microscopy confocal. In HeLa cells, both SDPR and PTRF were up-regulated by serum deprivation. We also found the N-terminal of SDPR protein was responsible for the interaction with PTRF. Overexpression of SDPR led to reduction of insulin induced ERK1/2 phosphorylation. However, the functional role of SDPR and its interaction with PTRF in insulin signaling pathway remains to be determined.