Porcine HMGCR Inhibits Porcine Circovirus Type 2 Infection by Directly Interacting with the Viral Proteins
Abstract
:1. Introduction
2. Materials and Methods
2.1. Virus and Cells
2.2. Drugs and Antibodies
2.3. Plasmid Construction
2.4. Virus Infection and Treatment
2.5. Real-Time PCR
2.6. MTS Assay
2.7. Confocal Fluorescence Microscopy
2.8. Coimmunoprecipitation (Co-IP)
2.9. Western Blot (WB)
2.10. Statistical Analysis
3. Results
3.1. PCV2 Infection Increases Phosphorylation of PP2A, AMPK and HMGCR
3.2. Inhibition of HMGCR by Lovastatin Has No Effect on the Activities of AMPK and PP2A during PCV2 Infection
3.3. PP2A Has Little Effect on PCV2 Infection and HMGCR Activity
3.4. HMGCR Activity Is Mainly Regulated by AMPK during PCV2 Infection
3.5. Construction of Recombinant Expression Plasmids
3.6. HMGCR Interacts with the Cap Protein of PCV2
3.7. HMGCR Interacts with the Rep Protein of PCV2
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Primer | Oligonucleotide Sequences (5′–3′) | Cloning Site | Size (bp) |
---|---|---|---|
Rep-F | acgcgtatgtacccatacgacgtaccagattacgctcccagcaaaaagaatggaagaag | Mlu I Age I | 945 |
Rep-R | cgaccggttcagtaatttatttcatatggaaattcag | ||
Cap-F | acgcgtatggagcagaagctgatctcagaggaggacctgacgtatccaaggaggcgtta | Mlu I Age I | 708 |
Cap-R | cgaccggtttaagggttaagtggggggtctt | ||
HMGCR-F | ggatccatgttgtcaagactcttccgaatgc | BamH I EcoR I | 2674 |
HMGCR-R | gaattctcaagctgccttcttagtgcaag |
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Ouyang, T.; Niu, G.; Zhang, Y.; Liu, X.; Zhang, X.; Zhang, S.; Geng, Y.; Pang, D.; Ouyang, H.; Ren, L. Porcine HMGCR Inhibits Porcine Circovirus Type 2 Infection by Directly Interacting with the Viral Proteins. Viruses 2019, 11, 544. https://doi.org/10.3390/v11060544
Ouyang T, Niu G, Zhang Y, Liu X, Zhang X, Zhang S, Geng Y, Pang D, Ouyang H, Ren L. Porcine HMGCR Inhibits Porcine Circovirus Type 2 Infection by Directly Interacting with the Viral Proteins. Viruses. 2019; 11(6):544. https://doi.org/10.3390/v11060544
Chicago/Turabian StyleOuyang, Ting, Guyu Niu, Yifang Zhang, Xiaohua Liu, Xinwei Zhang, Shiqi Zhang, Yulu Geng, Daxin Pang, Hongsheng Ouyang, and Linzhu Ren. 2019. "Porcine HMGCR Inhibits Porcine Circovirus Type 2 Infection by Directly Interacting with the Viral Proteins" Viruses 11, no. 6: 544. https://doi.org/10.3390/v11060544