Characterization of Nuclear Protein SP110c

• Main Authors: Mu-Chuan Yao, 姚木銓
• Other Authors: Bo-Shiun Yan
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/17642701186844300005

碩士 === 國立陽明大學 === 微生物及免疫學研究所 === 98 === It is estimated that approximately one-third of human population has been infected by virulent Mycobacterium tuberculosis (MTB). However, yet only about 10% of those infected develop tuberculosis. Therefore, the key of susceptibility to clinical tuberculosis is worthy to investigate. Genetic variation within host population is known to play a role in resistance and susceptibility to the pathogen. In previous studies, a gene,Ipr1 (Intracellular pathogen resistance 1), which mediates host innate immunity to MTB infection in mouse model, was identified within sst1 (supersusceptibility to tuberculosis 1) locus on mouse chromosome 1. In human, the closest homologue of predicted IPR1 protein is SP110b (41% of identity), which localizes to a region of human chromosome 2. Both IPR1 and human SP110b are not only similar in structure but also regulated by interferons. It’s suggested that they may have the same function in mouse and human. SP110 proteins have 3 major isoforms: SP110a, SP110b and SP110c). SP110c is the longest isoform, and SP110b is an alternative splicing isoform from SP110c. The alternative splicing makes different isoforms of SP110; however, whether these isoforms resulted from the alternative splicing have different roles and function is still unclear. In this study, we used dual-promoter lentiviral system to establish the SP110c-THP-1 stable clone, which can be induced to express eGFP-SP110c protein by doxycycline. The stability of eGFP-SP110c protein was increased after cells were differentiated by PMA treatment and remarkably increased after IFN-γ treatment. These results indicate that SP110c protein may be more stable in pro-inflammation environment. The proteins interacting with eGFP- SP110c were further characterized. To this end, the protein complex interacting with eGFP-SP110c was pull down by co-immunoprecipitation. We will identify the proteins interacting with SP110c by MS/MS analysis and compare the protein complex profile with that of SP110b.