This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Means, J. C.
Right arrow Articles by Clem, R. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Means, J. C.
Right arrow Articles by Clem, R. J.

 Previous Article  |  Next Article 

Journal of Virology, April 2003, p. 4481-4488, Vol. 77, No. 8
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.8.4481-4488.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Silencing of the Baculovirus Op-iap3 Gene by RNA Interference Reveals that It Is Required for Prevention of Apoptosis during Orgyia pseudotsugata M Nucleopolyhedrovirus Infection of Ld652Y Cells{dagger}

John C. Means, Israel Muro, and Rollie J. Clem*

Molecular, Cellular, and Developmental Biology Program, Division of Biology, Kansas State University, Manhattan, Kansas 66506

Received 13 November 2002/ Accepted 14 January 2003

The Op-iap3 gene from the baculovirus Orgyia pseudotsugata M nucleopolyhedrovirus (OpMNPV) inhibits apoptosis induced by a mutant of Autographa californica MNPV (AcMNPV) that lacks the antiapoptotic gene p35, as well as apoptosis induced by a wide range of other stimuli in both mammalian and insect cells. However, the role of Op-iap3 during OpMNPV infection has not been previously examined. To determine the function of the Op-IAP3 protein during OpMNPV infection, we used RNA interference (RNAi) to silence Op-iap3 expression during OpMNPV infection of Ld652Y cells. Infected cells treated with Op-iap3 double-stranded RNA (dsRNA) did not accumulate detectable Op-iap3 mRNA, confirming that the Op-iap3 gene was effectively silenced. Op-IAP3 protein was found to be a component of the budded virion; however, in OpMNPV-infected cells treated with Op-iap3 dsRNA, the Op-IAP3 protein that was introduced by the inoculum virus decreased to almost undetectable levels by 12 h after dsRNA addition. Apoptosis was observed in infected cells treated with Op-iap3 dsRNA beginning at 12 h, and by 48 h, almost all of the cells had undergone apoptosis. These results show for the first time that Op-IAP3 is necessary to prevent apoptosis during OpMNPV infection. In addition, our results demonstrate that the RNAi technique can be an effective tool for studying baculovirus gene function.

* Corresponding author. Mailing address: Division of Biology, 232 Ackert Hall, Kansas State University, Manhattan, KS 66506. Phone: (785) 532-3172. Fax: (785) 532-6653. E-mail: rclem{at}ksu.edu.

{dagger} Contribution no. 03-165-J from the Kansas Agricultural Experiment Station.

Journal of Virology, April 2003, p. 4481-4488, Vol. 77, No. 8
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.8.4481-4488.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Schultz, K. L. W., Friesen, P. D. (2009). Baculovirus DNA Replication-Specific Expression Factors Trigger Apoptosis and Shutoff of Host Protein Synthesis during Infection. J. Virol. 83: 11123-11132 [Abstract] [Full Text]  
  • Schultz, K. L. W., Wetter, J. A., Fiore, D. C., Friesen, P. D. (2009). Transactivator IE1 Is Required for Baculovirus Early Replication Events That Trigger Apoptosis in Permissive and Nonpermissive Cells. J. Virol. 83: 262-272 [Abstract] [Full Text]  
  • Yu, Q., Lin, T., Feng, G., Yang, K., Pang, Y. (2008). Functional analysis of the putative antiapoptotic genes, p49 and iap4, of Spodoptera litura nucleopolyhedrovirus with RNAi. J. Gen. Virol. 89: 1873-1880 [Abstract] [Full Text]  
  • Quadt, I., van Lent, J. W. M., Knebel-Morsdorf, D. (2007). Studies of the Silencing of Baculovirus DNA Binding Protein. J. Virol. 81: 6122-6127 [Abstract] [Full Text]  
  • Knebel-Morsdorf, D., Quadt, I., Li, Y., Montier, L., Guarino, L. A. (2006). Expression of Baculovirus Late and Very Late Genes Depends on LEF-4, a Component of the Viral RNA Polymerase Whose Guanyltransferase Function Is Essential.. J. Virol. 80: 4168-4173 [Abstract] [Full Text]  
  • Li, Q., Liston, P., Moyer, R. W. (2005). Functional Analysis of the Inhibitor of Apoptosis (iap) Gene Carried by the Entomopoxvirus of Amsacta moorei. J. Virol. 79: 2335-2345 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»