Interaction analyses between the nervous necrosis virus (NNV)coat protein and RNF2 proteins

• Main Authors: Sim-kun Ng, 吳嬋娟
• Other Authors: Hau-Ren Chen
• Format: Others
• Language: en_US
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/42445509446355565614

碩士 === 國立中正大學 === 生命科學系暨分子生物研究所暨生物醫學研究 === 97 === Viral nervous necrosis (VNN) is a worldwide disease among marine fishes and causes high mortalities at larval stage and considerable economic damage to the aquaculture industry. Nervous necrosis virus (NNV) is one of the major pathogens of VNN diseases and infects the larvae of orange-spotted grouper (Epinephelus coiodes) seriously. According to previous studies by Jimmy Kwang, the coat protein of NNV could enter the nucleus and trigger the process of apoptosis, implying that coat protein was not only a structural protein but also a potential protein involved in cellular genes regulation. To further study the functions of coat protein, yeast two-hybrid assay was used to screen for the NNV coat-associated proteins using the human fetal brain cDNA library. An alternative splicing form of human Ring finger protein 2 (hRNF2S) was identified. Surprisingly, NNV coat protein interacted with hRNF2S, but not with the full length hRNF2. In this thesis, yeast two-hybrid assay and GST-pull down assay were performed to further confirm the interaction and map the interaction domain between NNV coat and hRNF2 S / hRNF2 FL. NNV coat protein mainly interacts with the N-terminal region of hRNF2S, especially the RING finger domain of RNF2. Since NNV infects fishes naturally, the interaction between NNV coat and fish orange-spotted grouper (Osg) RNF2 was examined both in vitro and in vivo by yeast two-hybrid assay, GST pull-down assay, co-immunoprecipitation assay and immunoflurorescence assay. Thus, the biological function of this interaction will be further studied in the future.