title: Role and fate of SP100 protein in response to Rep-dependent nonviral integration system.
abstract: Previously, we studied an AAVS1 site-specific non-viral integration system with a Rep-donor plasmid and a plasmid containing adeno-associated virus integration element. Our earlier study focused on the plasmid vector itself, but the cellular response to the system was still unknown. SP100 is a member of the promyelocytic leukemia nuclear bodies. It is involved in many cellular processes such as transcriptional regulation and the cellular intrinsic immune response against viral infection. In this study, we revealed that SP100 inhibited the Rep-dependent nonviral integration. Conversely, transient expression of Rep78 increased the degradation of SP100. This degradation was inhibited by treatment with MG132, an inhibitor of the ubiquitin proteasome. SP100 and Rep78 are both located in the nucleolus, which provides the spatial possibility for their interaction. Rep78 was coimmunoprecipitated with the enhanced green fluorescent protein (EGFP)-SP100 fusion protein but not EGFP, which verified the interaction between Rep78 and SP100. These results have enriched our knowledge about the cellular protein SP100 and Rep-dependent nonviral integration. It may lead to an improvement in the application of Rep-related transgene integration method and in the selection of target cells.
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, 200433, People's Republic of China.
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Xue
authors.forename: Ran
authors.initials: R
authors.lastname: Wang
authors.forename: Yang-Bo
authors.initials: YB
authors.lastname: Yue
authors.forename: Jing-Lun
authors.initials: JL
authors.lastname: Xue
authors.forename: Jin-Zhong
authors.initials: JZ
authors.lastname: Chen
mesh_terms: D034961:Antigens, Nuclear / Q000378:metabolism*; D001324:Autoantigens / Q000378:metabolism*; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D010957:Plasmids; D011485:Protein Binding; D014764:Viral Proteins / Q000378:metabolism; D016662:Virus Integration
chemical_list: D034961:Antigens, Nuclear; D001324:Autoantigens; D004268:DNA-Binding Proteins; D014764:Viral Proteins; C066642:SP100 protein, human; C068734:rep proteins, Adeno-associated virus 2