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Chinese Journal of Biotechnology ; (12): 1985-1995, 2018.
Artigo em Chinês | WPRIM | ID: wpr-771410

RESUMO

Several putative transcription factor binding sites (TFBSs) exist in the PCV2 rep gene promoter. To explore if porcine circovirus type 2 (PCV2) could regulate the viral replication by using these TFBSs, we conducted electrophoretic mobility shift assay (EMSA), DNA-pull down and liquid chromatography-tandem mass spectrometric (LC-MS/MS) assays. EMSA confirmed the binding activity of the rep gene promoter with nuclear proteins of host cells. DNA-pull down and LC-MS/MS identified the porcine transcription factor AP-2δ (poTFAP2δ) could bind the PCV2 rep gene promoter. Dual-luciferase reporter assay, quantitative real-time PCR, Western blotting and indirect immunofluorescent assay demonstrated that poTFAP2δ could not only promote the activity of the rep gene promoter, but also enhance the transcription/translation activity of the rep/cap gene and the virus titer of PCV2 during the entire life cycle of PCV2 infection. This study revealed the molecular mechanism of PCV2 using host proteins to enhance the viral replication, provided a new perspective for studying the pathogenic mechanism of PCV2 from virus and host interactions, and provided a theoretical basis for developing highly effective PCV2 vaccines.


Assuntos
Animais , Linhagem Celular , Cromatografia Líquida , Infecções por Circoviridae , Circovirus , DNA Helicases , Diabetes Mellitus Tipo 2 , Regiões Promotoras Genéticas , Suínos , Espectrometria de Massas em Tandem , Fator de Transcrição AP-2 , Replicação Viral
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