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 Angulo, A.
Right arrow Articles by Messerle, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Angulo, A.
Right arrow Articles by Messerle, M.

 Previous Article  |  Next Article 

Journal of Virology, December 2000, p. 11129-11136, Vol. 74, No. 23
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The Major Immediate-Early Gene ie3 of Mouse Cytomegalovirus Is Essential for Viral Growth†

Ana Angulo,1,* Peter Ghazal,1 and Martin Messerle2,*

Department of Immunology and Molecular Biology, Division of Virology, The Scripps Research Institute, La Jolla, California 92037,1 and Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Genzentrum der Ludwig-Maximilians-Universität München, D-81377 Munich, Germany2

Received 9 June 2000/Accepted 30 August 2000

The significance of the major immediate-early gene ie3 of mouse cytomegalovirus (MCMV) and that of the corresponding ie2 gene of human cytomegalovirus to viral replication are not known. To investigate the function of the MCMV IE3 regulatory protein, we generated two different MCMV recombinants that contained a large deletion in the IE3 open reading frame (ORF). The mutant genomes were constructed by the bacterial artificial chromosome mutagenesis technique, and MCMV ie3 deletion mutants were reconstituted on a mouse fibroblast cell line that expresses the MCMV major immediate-early genes. The ie3 deletion mutants failed to replicate on normal mouse fibroblasts even when a high multiplicity of infection was used. The replication defect was rescued when the IE3 protein was provided in trans by a complementing cell line. A revertant virus in which the IE3 ORF was restored was able to replicate with wild-type kinetics in normal mouse fibroblasts, providing evidence that the defective growth phenotype of the ie3 mutants was due to disruption of the ie3 gene. To characterize the point of restriction in viral replication that is controlled by ie3, we analyzed the pattern of expression of selective early (beta ) and late (gamma ) genes. While we could detect transcripts for the immediate-early gene ie1 in cells infected with the ie3 mutants, we failed to detect transcripts for representative beta  and gamma  genes. These data demonstrate that the MCMV transactivator IE3 plays an indispensable role during viral replication in tissue culture, implicating a similar role for the human CMV ie2 gene product. To our knowledge, the ie3 deletion mutants represent the first MCMV recombinants isolated that contain a disruption of an essential gene.

* Corresponding authors. Mailing address for Martin Messerle: Max von Pettenkofer-Institut, Genzentrum, Feodor-Lynen-Strasse 25, D-81377 Munich, Germany. Phone: 49 89 2180 6850. Fax: 49 89 2180 6898. E-mail: messerle{at}lmb.uni-muenchen.de. Mailing address for Ana Angulo: Department of Immunology and Molecular Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. Phone: (858) 784-9933. Fax: (858) 784-9272. E-mail: angulo{at}scripps.edu.

dagger Publication 13327-IMM from The Scripps Research Institute.

Journal of Virology, December 2000, p. 11129-11136, Vol. 74, No. 23
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Kropp, K. A., Simon, C. O., Fink, A., Renzaho, A., Kuhnapfel, B., Podlech, J., Reddehase, M. J., Grzimek, N. K. A. (2009). Synergism between the components of the bipartite major immediate-early transcriptional enhancer of murine cytomegalovirus does not accelerate virus replication in cell culture and host tissues. J. Gen. Virol. 90: 2395-2401 [Abstract] [Full Text]  
  • Seckert, C. K., Renzaho, A., Tervo, H.-M., Krause, C., Deegen, P., Kuhnapfel, B., Reddehase, M. J., Grzimek, N. K. A. (2009). Liver Sinusoidal Endothelial Cells Are a Site of Murine Cytomegalovirus Latency and Reactivation. J. Virol. 83: 8869-8884 [Abstract] [Full Text]  
  • Wilhelmi, V., Simon, C. O., Podlech, J., Bohm, V., Daubner, T., Emde, S., Strand, D., Renzaho, A., Lemmermann, N. A. W., Seckert, C. K., Reddehase, M. J., Grzimek, N. K. A. (2008). Transactivation of Cellular Genes Involved in Nucleotide Metabolism by the Regulatory IE1 Protein of Murine Cytomegalovirus Is Not Critical for Viral Replicative Fitness in Quiescent Cells and Host Tissues. J. Virol. 82: 9900-9916 [Abstract] [Full Text]  
  • Wiebusch, L., Neuwirth, A., Grabenhenrich, L., Voigt, S., Hagemeier, C. (2008). Cell Cycle-Independent Expression of Immediate-Early Gene 3 Results in G1 and G2 Arrest in Murine Cytomegalovirus-Infected Cells. J. Virol. 82: 10188-10198 [Abstract] [Full Text]  
  • Buck, A. H., Santoyo-Lopez, J., Robertson, K. A., Kumar, D. S., Reczko, M., Ghazal, P. (2007). Discrete Clusters of Virus-Encoded MicroRNAs Are Associated with Complementary Strands of the Genome and the 7.2-Kilobase Stable Intron in Murine Cytomegalovirus. J. Virol. 81: 13761-13770 [Abstract] [Full Text]  
  • Kawasaki, H., Mocarski, E. S., Kosugi, I., Tsutsui, Y. (2007). Cyclosporine Inhibits Mouse Cytomegalovirus Infection via a Cyclophilin-Dependent Pathway Specifically in Neural Stem/Progenitor Cells. J. Virol. 81: 9013-9023 [Abstract] [Full Text]  
  • Simon, C. O., Kuhnapfel, B., Reddehase, M. J., Grzimek, N. K. A. (2007). Murine Cytomegalovirus Major Immediate-Early Enhancer Region Operating as a Genetic Switch in Bidirectional Gene Pair Transcription. J. Virol. 81: 7805-7810 [Abstract] [Full Text]  
  • Simon, C. O., Holtappels, R., Tervo, H.-M., Bohm, V., Daubner, T., Oehrlein-Karpi, S. A., Kuhnapfel, B., Renzaho, A., Strand, D., Podlech, J., Reddehase, M. J., Grzimek, N. K. A. (2006). CD8 T Cells Control Cytomegalovirus Latency by Epitope-Specific Sensing of Transcriptional Reactivation. J. Virol. 80: 10436-10456 [Abstract] [Full Text]  
  • Pinto, A. K., Munks, M. W., Koszinowski, U. H., Hill, A. B. (2006). Coordinated Function of Murine Cytomegalovirus Genes Completely Inhibits CTL Lysis.. J. Immunol. 177: 3225-3234 [Abstract] [Full Text]  
  • Tripathy, S. K., Smith, H. R. C., Holroyd, E. A., Pingel, J. T., Yokoyama, W. M. (2006). Expression of m157, a Murine Cytomegalovirus-Encoded Putative Major Histocompatibility Class I (MHC-I)-Like Protein, Is Independent of Viral Regulation of Host MHC-I. J. Virol. 80: 545-550 [Abstract] [Full Text]  
  • Cicin-Sain, L., Podlech, J., Messerle, M., Reddehase, M. J., Koszinowski, U. H. (2005). Frequent Coinfection of Cells Explains Functional In Vivo Complementation between Cytomegalovirus Variants in the Multiply Infected Host. J. Virol. 79: 9492-9502 [Abstract] [Full Text]  
  • Ghazal, P., Visser, A. E., Gustems, M., Garcia, R., Borst, E. M., Sullivan, K., Messerle, M., Angulo, A. (2005). Elimination of ie1 Significantly Attenuates Murine Cytomegalovirus Virulence but Does Not Alter Replicative Capacity in Cell Culture. J. Virol. 79: 7182-7194 [Abstract] [Full Text]  
  • Tang, Q., Li, L., Maul, G. G. (2005). Mouse Cytomegalovirus Early M112/113 Proteins Control the Repressive Effect of IE3 on the Major Immediate-Early Promoter. J. Virol. 79: 257-263 [Abstract] [Full Text]  
  • Simon, C. O., Seckert, C. K., Dreis, D., Reddehase, M. J., Grzimek, N. K. A. (2005). Role for Tumor Necrosis Factor Alpha in Murine Cytomegalovirus Transcriptional Reactivation in Latently Infected Lungs. J. Virol. 79: 326-340 [Abstract] [Full Text]  
  • Lembo, D., Donalisio, M., Hofer, A., Cornaglia, M., Brune, W., Koszinowski, U., Thelander, L., Landolfo, S. (2004). The Ribonucleotide Reductase R1 Homolog of Murine Cytomegalovirus Is Not a Functional Enzyme Subunit but Is Required for Pathogenesis. J. Virol. 78: 4278-4288 [Abstract] [Full Text]  
  • Benedict, C. A., Angulo, A., Patterson, G., Ha, S., Huang, H., Messerle, M., Ware, C. F., Ghazal, P. (2004). Neutrality of the Canonical NF-{kappa}B-Dependent Pathway for Human and Murine Cytomegalovirus Transcription and Replication In Vitro. J. Virol. 78: 741-750 [Abstract] [Full Text]  
  • Wang, X., Messerle, M., Sapinoro, R., Santos, K., Hocknell, P. K., Jin, X., Dewhurst, S. (2003). Murine Cytomegalovirus Abortively Infects Human Dendritic Cells, Leading to Expression and Presentation of Virally Vectored Genes. J. Virol. 77: 7182-7192 [Abstract] [Full Text]  
  • Isomura, H., Stinski, M. F. (2003). The Human Cytomegalovirus Major Immediate-Early Enhancer Determines the Efficiency of Immediate-Early Gene Transcription and Viral Replication in Permissive Cells at Low Multiplicity of Infection. J. Virol. 77: 3602-3614 [Abstract] [Full Text]  
  • Ghazal, P., Messerle, M., Osborn, K., Angulo, A. (2003). An Essential Role of the Enhancer for Murine Cytomegalovirus In Vivo Growth and Pathogenesis. J. Virol. 77: 3217-3228 [Abstract] [Full Text]  
  • Kercher, L., Mitchell, B. M. (2002). Persisting Murine Cytomegalovirus Can Reactivate and Has Unique Transcriptional Activity in Ocular Tissue. J. Virol. 76: 9165-9175 [Abstract] [Full Text]  
  • Gawn, J. M., Greaves, R. F. (2002). Absence of IE1 p72 Protein Function during Low-Multiplicity Infection by Human Cytomegalovirus Results in a Broad Block to Viral Delayed-Early Gene Expression. J. Virol. 76: 4441-4455 [Abstract] [Full Text]  
  • Heider, J. A., Bresnahan, W. A., Shenk, T. E. (2002). Construction of a rationally designed human cytomegalovirus variant encoding a temperature-sensitive immediate-early 2 protein. Proc. Natl. Acad. Sci. USA 10.1073/pnas.052710599v1 [Abstract] [Full Text]  
  • Meier, J. L., Keller, M. J., McCoy, J. J. (2002). Requirement of Multiple cis-Acting Elements in the Human Cytomegalovirus Major Immediate-Early Distal Enhancer for Viral Gene Expression and Replication. J. Virol. 76: 313-326 [Abstract] [Full Text]  
  • Sandford, G. R., Brock, L. E., Voigt, S., Forester, C. M., Burns, W. H. (2001). Rat Cytomegalovirus Major Immediate-Early Enhancer Switching Results in Altered Growth Characteristics. J. Virol. 75: 5076-5083 [Abstract] [Full Text]  
  • Grzimek, N. K. A., Dreis, D., Schmalz, S., Reddehase, M. J. (2001). Random, Asynchronous, and Asymmetric Transcriptional Activity of Enhancer-Flanking Major Immediate-Early Genes ie1/3 and ie2 during Murine Cytomegalovirus Latency in the Lungs. J. Virol. 75: 2692-2705 [Abstract] [Full Text]  
  • Holtappels, R., Pahl-Seibert, M.-F., Thomas, D., Reddehase, M. J. (2000). Enrichment of Immediate-Early 1 (m123/pp89) Peptide-Specific CD8 T Cells in a Pulmonary CD62Llo Memory-Effector Cell Pool during Latent Murine Cytomegalovirus Infection of the Lungs. J. Virol. 74: 11495-11503 [Abstract] [Full Text]  
  • Heider, J. A., Bresnahan, W. A., Shenk, T. E. (2002). Construction of a rationally designed human cytomegalovirus variant encoding a temperature-sensitive immediate-early 2 protein. Proc. Natl. Acad. Sci. USA 99: 3141-3146 [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»