A Novel Role for PX, a Structural Protein of Fowl Adenovirus Serotype 4 (FAdV4), as an Apoptosis-Inducer in Leghorn Male Hepatocellular Cell
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Virus
2.2. Reagents, Chemicals, and Antibodies
2.3. Constructions of Recombinant Plasmids
2.4. Apoptosis Assay
2.5. Western Blot Analysis
2.6. Confocal Laser Scanning Microscopy Assays
2.7. Measurement of FAdV4 Growth in LMH Cells
2.8. Statistical Analysis
3. Results
3.1. FAdV4 Induced Apoptosis in LMH Cells
3.2. The PX Protein Plays a Major Role in FAdV4-Induced Apoptosis in LMH Cells
3.3. Localization of PX Protein in the Nucleus is Required for PX-Induced Apoptosis
3.4. Amino Acids 11 and 129 Alanines of PX Are Crucial to PX-Induced Apoptosis
3.5. Inhibition of FAdV4-Induced Apoptosis by Inhibitors Restricted Viral Replication
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Chandra, R.; Shukla, S.K.; Kumar, M. The hydropericardium syndrome and inclusion body hepatitis in domestic fowl. Trop. Anim. Health Prod. 2000, 32, 99–111. [Google Scholar] [CrossRef]
- Griffin, B.D.; Nagy, E. Coding potential and transcript analysis of fowl adenovirus 4: Insight into upstream ORFs as common sequence features in adenoviral transcripts. J. Gen. Virol. 2011, 92, 1260–1272. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Y.; Liu, R.; Tian, K.; Wang, Z.; Yang, X.; Gao, D.; Zhang, Y.; Fu, J.; Wang, H.; Zhao, J. Fiber2 and hexon genes are closely associated with the virulence of the emerging and highly pathogenic fowl adenovirus 4. Emerg. Microbes Infect. 2018, 7, 199. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pan, Q.; Wang, J.; Gao, Y.; Cui, H.; Liu, C.; Qi, X.; Zhang, Y.; Wang, Y.; Wang, X. The Natural Large Genomic Deletion Is Unrelated to the Increased Virulence of the Novel Genotype Fowl Adenovirus 4 Recently Emerged in China. Viruses 2018, 10, 494. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lasaro, M.O.; Ertl, H.C. New insights on adenovirus as vaccine vectors. Mol. Ther. 2009, 17, 1333–1339. [Google Scholar] [CrossRef] [PubMed]
- Lichtenstein, D.L.; Toth, K.; Doronin, K.; Tollefson, A.E.; Wold, W.S. Functions and mechanisms of action of the adenovirus E3 proteins. Int. Rev. Immunol. 2004, 23, 75–111. [Google Scholar] [CrossRef] [PubMed]
- Sharma, S.; Andersson, A. Adenovirus E3 proteins help tumors to evade innate and adaptive immune responses. Cancer Biol. Ther. 2009, 8, 1133–1135. [Google Scholar] [CrossRef] [Green Version]
- Everett, R.D.; Chiocca, S.; Orr, A. The chicken adenovirus Gam1 protein, an inhibitor of the sumoylation pathway, partially complements ICP0-null mutant herpes simplex virus 1. J. Virol. 2014, 88, 5873–5876. [Google Scholar] [CrossRef] [Green Version]
- Chiocca, S.; Baker, A.; Cotten, M. Identification of a novel antiapoptotic protein, GAM-1, encoded by the CELO adenovirus. J. Virol. 1997, 71, 3168–3177. [Google Scholar] [CrossRef] [Green Version]
- Lee, T.W.; Lawrence, F.J.; Dauksaite, V.; Akusjarvi, G.; Blair, G.E.; Matthews, D.A. Precursor of human adenovirus core polypeptide Mu targets the nucleolus and modulates the expression of E2 proteins. J. Gen. Virol. 2004, 85, 185–196. [Google Scholar] [CrossRef]
- Martin-Gonzalez, N.; Hernando-Perez, M.; Condezo, G.N.; Perez-Illana, M.; Siber, A.; Reguera, D.; Ostapchuk, P.; Hearing, P.; San Martin, C.; de Pablo, P.J. Adenovirus major core protein condenses DNA in clusters and bundles, modulating genome release and capsid internal pressure. Nucleic Acids Res. 2019, 47, 9231–9242. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Puntener, D.; Engelke, M.F.; Ruzsics, Z.; Strunze, S.; Wilhelm, C.; Greber, U.F. Stepwise loss of fluorescent core protein V from human adenovirus during entry into cells. J. Virol. 2011, 85, 481–496. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Niu, Y.; Sun, Q.; Zhang, G.; Liu, X.; Shang, Y.; Xiao, Y.; Liu, S. Fowl adenovirus serotype 4-induced apoptosis, autophagy, and a severe inflammatory response in liver. Vet. Microbiol. 2018, 223, 34–41. [Google Scholar] [CrossRef] [PubMed]
- Reed, L.J.; Muench, H. A simple method of estimating fifty percent endpoints. Am. J. Hyg. 1938, 27, 493–497. [Google Scholar]
- Efthymiadis, A.; Shao, H.; Hubner, S.; Jans, D.A. Kinetic characterization of the human retinoblastoma protein bipartite nuclear localization sequence (NLS) in vivo and in vitro. A comparison with the SV40 large T-antigen NLS. J. Biol. Chem. 1997, 272, 22134–22139. [Google Scholar] [CrossRef] [Green Version]
- Duan, X.; Zhao, M.; Wang, Y.; Li, X.; Cao, H.; Zheng, S.J. Epigenetic Upregulation of Chicken MicroRNA-16-5p Expression in DF-1 Cells following Infection with Infectious Bursal Disease Virus (IBDV) Enhances IBDV-induced Apoptosis and Viral Replication. J. Virol. 2019, 94, e01724-01719. [Google Scholar] [CrossRef]
- Li, Z.; Wang, Y.; Xue, Y.; Li, X.; Cao, H.; Zheng, S.J. Critical role for voltage-dependent anion channel 2 in infectious bursal disease virus-induced apoptosis in host cells via interaction with VP5. J. Virol. 2012, 86, 1328–1338. [Google Scholar] [CrossRef] [Green Version]
- Jaffery, M.S. A treatise on Angara disease (hydropericardium-pulmonary oedema-hepatonephritis syndrome). J. Pakistan Vet. Med. Assoc. 1988, 34, 1–33. [Google Scholar]
- Cheema, A.E.; Ahmad, J.; Afzal, M. An adenovirus infection of poultry in Pakistan. Revue Sci. Tech. 1989, 8, 789–795. [Google Scholar] [CrossRef]
- Hess, M.; Raue, R.; Prusas, C. Epidemiological studies on fowl adenoviruses isolated from cases of infectious hydropericardium. Avian Pathol. 1999, 28, 433–439. [Google Scholar] [CrossRef]
- Kim, J.N.; Byun, S.H.; Kim, M.J.; Kim, J.; Sung, H.W.; Mo, I.P. Outbreaks of hydropericardium syndrome and molecular characterization of Korean fowl adenoviral isolates. Avian Dis. 2008, 52, 526–530. [Google Scholar] [CrossRef] [PubMed]
- Asrani, R.K.; Gupta, V.K.; Sharma, S.K.; Singh, S.P.; Katoch, R.C. Hydropericardium-hepatopathy syndrome in Asian poultry. Vet. Rec. 1997, 141, 271–273. [Google Scholar] [CrossRef] [PubMed]
- Shen, Z.; Xiang, B.; Li, S.; Ren, X.; Hong, Y.; Liao, J.; Yu, D.; Ren, T.; Liao, M.; Xu, C. Genetic characterization of fowl adenovirus serotype 4 isolates in Southern China reveals potential cross-species transmission. Infect. Genet. Evol. 2019, 75, 103928. [Google Scholar] [CrossRef] [PubMed]
- Chiocca, S.; Kurzbauer, R.; Schaffner, G.; Baker, A.; Mautner, V.; Cotten, M. The complete DNA sequence and genomic organization of the avian adenovirus CELO. J. Virol. 1996, 70, 2939–2949. [Google Scholar] [CrossRef] [Green Version]
- Harrison, S.C. Virology. Looking inside adenovirus. Science 2010, 329, 1026–1027. [Google Scholar] [CrossRef]
- Gao, J.; Zhao, M.; Duan, X.; Wang, Y.; Cao, H.; Li, X.; Zheng, S.J. Requirement of Cellular Protein CCT7 for the Replication of Fowl Adenovirus Serotype 4 (FAdV-4) in Leghorn Male Hepatocellular Cells Via Interaction with the Viral Hexon Protein. Viruses 2019, 11, 107. [Google Scholar] [CrossRef] [Green Version]
- Li, L.; Wang, J.; Chen, P.; Zhang, S.; Sun, J.; Yuan, W. Pathogenicity and molecular characterization of a fowl adenovirus 4 isolated from chicken associated with IBH and HPS in China. BMC Vet. Res. 2018, 14, 400. [Google Scholar] [CrossRef] [Green Version]
- San Martin, C. Latest insights on adenovirus structure and assembly. Viruses 2012, 4, 847–877. [Google Scholar] [CrossRef] [Green Version]
- Hong, S.S.; Szolajska, E.; Schoehn, G.; Franqueville, L.; Myhre, S.; Lindholm, L.; Ruigrok, R.W.; Boulanger, P.; Chroboczek, J. The 100K-chaperone protein from adenovirus serotype 2 (Subgroup C) assists in trimerization and nuclear localization of hexons from subgroups C and B adenoviruses. J. Mol. Biol. 2005, 352, 125–138. [Google Scholar] [CrossRef]
- Hong, S.S.; Gay, B.; Karayan, L.; Dabauvalle, M.C.; Boulanger, P. Cellular uptake and nuclear delivery of recombinant adenovirus penton base. Virology 1999, 262, 163–177. [Google Scholar] [CrossRef] [Green Version]
- Hammond, C.M.; Stromme, C.B.; Huang, H.; Patel, D.J.; Groth, A. Histone chaperone networks shaping chromatin function. Nat. Rev. Mol. Cell Biol. 2017, 18, 141–158. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Terry, L.J.; Shows, E.B.; Wente, S.R. Crossing the Nuclear Envelope Hierarchical Regulation of Nucleocytoplasmic Transport. Science 2007, 318, 1412–1416. [Google Scholar] [CrossRef] [PubMed]
- Liu, Y.; Wan, W.; Gao, D.; Li, Y.; Yang, X.; Liu, H.; Yao, H.; Chen, L.; Wang, C.; Zhao, J. Genetic characterization of novel fowl aviadenovirus 4 isolates from outbreaks of hepatitis-hydropericardium syndrome in broiler chickens in China. Emerg. Microbes Infect. 2016, 5, e117. [Google Scholar] [CrossRef] [PubMed]
- Zou, X.H.; Bi, Z.X.; Guo, X.J.; Zhang, Z.; Zhao, Y.; Wang, M.; Zhu, Y.L.; Jie, H.Y.; Yu, Y.; Hung, T.; et al. DNA assembly technique simplifies the construction of infectious clone of fowl adenovirus. J. Virol. Methods 2018, 257, 85–92. [Google Scholar] [CrossRef] [PubMed]
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Zhao, M.; Duan, X.; Wang, Y.; Gao, L.; Cao, H.; Li, X.; Zheng, S.J. A Novel Role for PX, a Structural Protein of Fowl Adenovirus Serotype 4 (FAdV4), as an Apoptosis-Inducer in Leghorn Male Hepatocellular Cell. Viruses 2020, 12, 228. https://doi.org/10.3390/v12020228
Zhao M, Duan X, Wang Y, Gao L, Cao H, Li X, Zheng SJ. A Novel Role for PX, a Structural Protein of Fowl Adenovirus Serotype 4 (FAdV4), as an Apoptosis-Inducer in Leghorn Male Hepatocellular Cell. Viruses. 2020; 12(2):228. https://doi.org/10.3390/v12020228
Chicago/Turabian StyleZhao, Mingliang, Xueyan Duan, Yongqiang Wang, Li Gao, Hong Cao, Xiaoqi Li, and Shijun J. Zheng. 2020. "A Novel Role for PX, a Structural Protein of Fowl Adenovirus Serotype 4 (FAdV4), as an Apoptosis-Inducer in Leghorn Male Hepatocellular Cell" Viruses 12, no. 2: 228. https://doi.org/10.3390/v12020228
APA StyleZhao, M., Duan, X., Wang, Y., Gao, L., Cao, H., Li, X., & Zheng, S. J. (2020). A Novel Role for PX, a Structural Protein of Fowl Adenovirus Serotype 4 (FAdV4), as an Apoptosis-Inducer in Leghorn Male Hepatocellular Cell. Viruses, 12(2), 228. https://doi.org/10.3390/v12020228