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Sheng Wu Gong Cheng Xue Bao. 2018 Dec 25;34(12):1985-1995. doi: 10.13345/j.cjb.180069.

[Porcine transcription factor AP-2δ promotes porcine circovirus type 2 replication through enhancing the activity of the rep gene promoter].

[Article in Chinese; Abstract available in Chinese from the publisher]

Author information

1
Institute of Immunology, Nanjing Agricultural University, Nanjing 210095, Jiangsu, China.
2
Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, Zhejiang, China.

Abstract

in English, Chinese

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.

KEYWORDS:

porcine circovirus type 2; porcine transcription factor AP-2δ; rep gene promoter; transcription factor binding sites

PMID:
30584709
DOI:
10.13345/j.cjb.180069
[Indexed for MEDLINE]
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