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J OURNAL OF V IROLOGY,June2009,p.6039–6047Vol.83,No.12 0022-538X/09/$08.00?0doi:10.1128/JVI.00135-09

Copyright?2009,American Society for Microbiology.All Rights Reserved.

JNK and p38Mitogen-Activated Protein Kinase Pathways Contribute to Porcine Circovirus Type2Infection?

Li Wei,1Zhongwu Zhu,2Jing Wang,1and Jue Liu1*

Institute of Animal Husbandry and Veterinary Medicine,Beijing Municipal Academy of Agriculture and Forestry Sciences, No.9Shuguang Garden Central Road,Haidian District,Beijing100097,1and Hunan Entry-Exit Inspection and Quarantine Bureau,No.188Xiangfu Road,Changsha410003,2People’s Republic of China

Received21January2009/Accepted23March2009

Infection with a wide variety of viruses often perturbs host cell signaling pathways including the Jun

NH

2-terminal kinase/stress-activated kinase(JNK/SAPK)and the p38mitogen-activated protein kinase(p38/

MAPK),which are important components of cellular signal transduction pathways.The present study dem-onstrated for the?rst time that porcine circovirus type2(PCV2),which is the primary causative agent of an emerging swine disease,postweaning multisystemic wasting syndrome,can activate JNK1/2and p38MAPK pathways in PCV2-infected PK15cells.However,PCV2at an early stage of infection,as well as UV-irradiated PCV2,failed to activate these two MAPK families,which demonstrated that PCV2replication was necessary for their activation.We further found that PCV2activated the phosphorylation of JNK1/2and p38MAPK downstream targets c-Jun and ATF-2with virus replication in the cultured cells.The roles of these kinases in PCV2infection were further evaluated using speci?c inhibitors:the JNK inhibitor1for JNK1/2and SB202190 for p38.Inhibition of JNK1/2and p38kinases by these speci?c inhibitors did result in signi?cant reduction of PCV2viral mRNA transcription and protein synthesis,viral progeny release,and blockage of PCV2-induced apoptotic caspase-3activation in the infected cells.Taken together,these data suggest that JNK/SAPK and p38 MAPK pathways play important roles in the PCV2replication and contribute to virus-mediated changes in host cells.

Porcine circovirus(PCV)is classi?ed in the genus Circovirus of the family Circoviridae(52).PCV was?rst discovered in 1974as a contaminant of a continuous porcine kidney cell line (PK15)(49).Two genotypes of PCV have been identi?ed.PCV type1(PCV1)is known to be nonpathogenic to pigs(1).A high prevalence of anti-PCV1antibodies has been detected in the swine population,but no disease is correlated with PCV1 infection(33,50).In contrast,infection with PCV2occurs in all swine-producing areas of the world,and related PCV2-associ-ated diseases are increasingly recognized as serious threats to global pig production(2,7,11,13,34,39,43).A primary manifestation of PCV2infection is postweaning multisystemic wasting syndrome(PMWS),which appears in pigs aged5to18 weeks and is clinically characterized by fever,wasting or un-thriftiness,respiratory distress,enlarged lymph nodes and,oc-casionally,jaundice and diarrhea(8,19,43).Mortality rates may vary from1to2%up to30%in complicated cases when coinfections with porcine reproductive and respiratory syn-drome virus,porcine parvovirus,or Mycoplasma hyopneu-moniae(12).Microscopic lesions are characterized by lympho-cyte depletion of follicular and interfollicular areas together with macrophage in?ltration of lymphoid tissues in PMWS-affected pigs.Several lines of?eld and experimental evidence have suggested that severely PMWS affected pigs may develop immunosuppression(44).

PCV genome is a circular single-stranded DNA molecule of ?1.7kb.Two major open reading frames(ORFs)have been recognized for PCV:ORF1,called the rep gene,which encodes a protein of35.7kDa involved in virus replication(35),and ORF2,called the cap gene,which encodes the major immu-nogenic capsid protein of27.8kDa(5,38).In addition to the replicase ORF1and the capsid protein ORF2,a novel protein, ORF3,has been detected in PCV2productive infection and is not essential for PCV2replication in the cultured cells but is involved in viral pathogenesis via an apoptotic function(30, 31).In a recent report,we demonstrated that PCV2infection induces NF-?B activation in the cultured cells and further found the role of NF-?B activation in PCV2replication and PCV2-induced apoptotic caspase activity(56).However, whether other signaling pathways may also contribute to PCV2 infection in the cultured cells remains unclear.

Mitogen-activated protein kinases(MAPKs),including ex-tracellular signal-regulated kinases(ERK1/2),c-Jun NH

2

-ter-minal kinase/stress-activated protein kinase(JNK/SAPK),and p38MAPK,are central components of signal transduction pathways in the regulation of cell proliferation and differenti-ation,cytokine production,and apoptosis(17).ERK1/2is pri-marily activated by growth factors,cytokines,and phagocytosis, whereas JNK and p38are potently induced by proin?am-matory cytokines,bacterial endotoxins,and environmental stresses(17,26).Although there is coordinated regulation of JNK/SAPK and p38MAPK,they have corresponding downstream targets.Activated JNK/SAPK and p38MAPK can phosphorylate numerous substrates,including a variety

*Corresponding author.Mailing address:Institute of Animal Hus-bandry and Veterinary Medicine,Beijing Municipal,Academy of Agri-culture and Forestry Sciences,No.9Shuguang Garden Central Road, Haidian District,Beijing100097,People’s Republic of China.Phone:

86-10-51503548.Fax:86-10-88433070.E-mail:liujue@http://www.doczj.com/doc/45b9ec6cbcd126fff7050b81.html.

?Published ahead of print on1April2009.

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