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 Ahn, J.-H.
Right arrow Articles by Hayward, G. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ahn, J.-H.
Right arrow Articles by Hayward, G. S.

 Previous Article  |  Next Article 

Journal of Virology, December 1999, p. 10458-10471, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Human Cytomegalovirus IE2 and UL112-113 Proteins Accumulate in Viral DNA Replication Compartments That Initiate from the Periphery of Promyelocytic Leukemia Protein-Associated Nuclear Bodies (PODs or ND10)

Jin-Hyun Ahn,1 Won-Jong Jang,1 and Gary S. Hayward1,2,*

Molecular Virology Laboratories, Departments of Pharmacology and Molecular Sciences1 and Oncology,2 Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

Received 6 July 1999/Accepted 24 August 1999

During human cytomegalovirus (HCMV) infection, the periphery of promyelocytic leukemia protein (PML)-associated nuclear bodies (also known as PML oncogenic domains [PODs] or ND10) are sites for both input viral genome deposition and immediate-early (IE) gene transcription. At very early times after infection, the IE1 protein localizes to and subsequently disrupts PODs, whereas the IE2 protein localizes within or adjacent to PODs. This process appears to be required for efficient viral gene expression and DNA replication. We have investigated the initiation of viral DNA replication compartment formation by studying the localization of viral IE proteins, DNA replication proteins, and the PML protein during productive infection. Localization of IE2 adjacent to PODs between 2 and 6 h after infection was confirmed by confocal microscopy of human fibroblasts (HF cells) infected with both wild-type HCMV(Towne) and with an IE1-deletion mutant HCMV(CR208) that fails to disrupt PODs. In HCMV(Towne)-infected HF cells at 24 to 48 h, IE2 also accumulated in newly formed viral DNA replication compartments containing the polymerase processivity factor (UL44), the single-stranded DNA binding protein (SSB; UL57), the UL112-113 accessory protein, and newly incorporated bromodeoxyuridine (BrdU). Double labeling of the HCMV(CR208)-infected HF cells demonstrated that formation of viral DNA replication compartments initiates within granular structures that bud from the periphery of some of the PODs and subsequently coalesce into larger structures that are flanked by PODs. In transient DNA transfection assays, both the N terminus (codons 136 to 290) and the C terminus (codons 379 to 579) of IE2 exon 5, but not the central region between them, were found to be necessary for both the punctate distribution of IE2 and its association with PODs. Like IE2, the UL112-113 accessory replication protein was also distributed in a POD-associated pattern in both DNA-transfected and virus-infected cells beginning at 6 h. Furthermore, when all six replication core machinery proteins (polymerase complex, SSB, and helicase-primase complex) were expressed together in the presence of UL112-113, they also accumulated at POD-associated sites, suggesting that the UL112-113 protein (but not IE2) may play a role in recruitment of viral replication fork proteins into the periphery of PODs. These results show that (i) subsequent to accumulating at the periphery of PODs, IE2 is incorporated together with the core proteins into viral DNA replication compartments that initiate from the periphery of PODs and then grow to fill the space between groups of PODs, and (ii) the UL112-113 protein appears to have a key role in assembling and recruiting the core replication machinery proteins in the initial stages of viral replication compartment formation.

* Corresponding author. Mailing address: Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, 725 N. Wolfe St., Baltimore, MD 21205. Phone: (410) 955-8684. Fax: (410) 955-8685. E-mail: ghayward{at}jhmi.edu.

Journal of Virology, December 1999, p. 10458-10471, Vol. 73, No. 12
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Kapasi, A. J., Clark, C. L., Tran, K., Spector, D. H. (2009). Recruitment of cdk9 to the Immediate-Early Viral Transcriptosomes during Human Cytomegalovirus Infection Requires Efficient Binding to Cyclin T1, a Threshold Level of IE2 86, and Active Transcription. J. Virol. 83: 5904-5917 [Abstract] [Full Text]  
  • Wells, R., Stensland, L., Vieira, J. (2009). The Human Cytomegalovirus UL112-113 Locus Can Activate the Full Kaposi's Sarcoma-Associated Herpesvirus Lytic Replication Cycle. J. Virol. 83: 4695-4699 [Abstract] [Full Text]  
  • Qian, Z., Xuan, B., Hong, T. T., Yu, D. (2008). The Full-Length Protein Encoded by Human Cytomegalovirus Gene UL117 Is Required for the Proper Maturation of Viral Replication Compartments. J. Virol. 82: 3452-3465 [Abstract] [Full Text]  
  • Kofod-Olsen, E., Ross-Hansen, K., Mikkelsen, J. G., Hollsberg, P. (2008). Human herpesvirus 6B U19 protein is a PML-regulated transcriptional activator that localizes to nuclear foci in a PML-independent manner. J. Gen. Virol. 89: 106-116 [Abstract] [Full Text]  
  • Kapasi, A. J., Spector, D. H. (2008). Inhibition of the Cyclin-Dependent Kinases at the Beginning of Human Cytomegalovirus Infection Specifically Alters the Levels and Localization of the RNA Polymerase II Carboxyl-Terminal Domain Kinases cdk9 and cdk7 at the Viral Transcriptosome. J. Virol. 82: 394-407 [Abstract] [Full Text]  
  • Park, J.-J., Kim, Y.-E., Pham, H. T., Kim, E. T., Chung, Y.-H., Ahn, J.-H. (2007). Functional interaction of the human cytomegalovirus IE2 protein with histone deacetylase 2 in infected human fibroblasts. J. Gen. Virol. 88: 3214-3223 [Abstract] [Full Text]  
  • Sourvinos, G., Tavalai, N., Berndt, A., Spandidos, D. A., Stamminger, T. (2007). Recruitment of Human Cytomegalovirus Immediate-Early 2 Protein onto Parental Viral Genomes in Association with ND10 in Live-Infected Cells. J. Virol. 81: 10123-10136 [Abstract] [Full Text]  
  • Tomoiu, A., Gravel, A., Tanguay, R. M., Flamand, L. (2006). Functional Interaction between Human Herpesvirus 6 Immediate-Early 2 Protein and Ubiquitin-Conjugating Enzyme 9 in the Absence of Sumoylation.. J. Virol. 80: 10218-10228 [Abstract] [Full Text]  
  • Tavalai, N., Papior, P., Rechter, S., Leis, M., Stamminger, T. (2006). Evidence for a Role of the Cellular ND10 Protein PML in Mediating Intrinsic Immunity against Human Cytomegalovirus Infections.. J. Virol. 80: 8006-8018 [Abstract] [Full Text]  
  • Park, M.-Y., Kim, Y.-E., Seo, M.-R., Lee, J.-R., Lee, C. H., Ahn, J.-H. (2006). Interactions among Four Proteins Encoded by the Human Cytomegalovirus UL112-113 Region Regulate Their Intranuclear Targeting and the Recruitment of UL44 to Prereplication Foci. J. Virol. 80: 2718-2727 [Abstract] [Full Text]  
  • Tamrakar, S., Kapasi, A. J., Spector, D. H. (2005). Human Cytomegalovirus Infection Induces Specific Hyperphosphorylation of the Carboxyl-Terminal Domain of the Large Subunit of RNA Polymerase II That Is Associated with Changes in the Abundance, Activity, and Localization of cdk9 and cdk7. J. Virol. 79: 15477-15493 [Abstract] [Full Text]  
  • Castillo, J. P., Frame, F. M., Rogoff, H. A., Pickering, M. T., Yurochko, A. D., Kowalik, T. F. (2005). Human Cytomegalovirus IE1-72 Activates Ataxia Telangiectasia Mutated Kinase and a p53/p21-Mediated Growth Arrest Response. J. Virol. 79: 11467-11475 [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]  
  • Isomura, H., Tsurumi, T., Stinski, M. F. (2004). Role of the Proximal Enhancer of the Major Immediate-Early Promoter in Human Cytomegalovirus Replication. J. Virol. 78: 12788-12799 [Abstract] [Full Text]  
  • Xu, Y., Cei, S. A., Huete, A. R., Pari, G. S. (2004). Human Cytomegalovirus UL84 Insertion Mutant Defective for Viral DNA Synthesis and Growth. J. Virol. 78: 10360-10369 [Abstract] [Full Text]  
  • Day, P. M., Baker, C. C., Lowy, D. R., Schiller, J. T. (2004). Establishment of papillomavirus infection is enhanced by promyelocytic leukemia protein (PML) expression. Proc. Natl. Acad. Sci. USA 101: 14252-14257 [Abstract] [Full Text]  
  • Lee, H.-R., Ahn, J.-H. (2004). Sumoylation of the major immediate-early IE2 protein of human cytomegalovirus Towne strain is not required for virus growth in cultured human fibroblasts. J. Gen. Virol. 85: 2149-2154 [Abstract] [Full Text]  
  • Lee, H.-R., Kim, D.-J., Lee, J.-M., Choi, C. Y., Ahn, B.-Y., Hayward, G. S., Ahn, J.-H. (2004). Ability of the Human Cytomegalovirus IE1 Protein To Modulate Sumoylation of PML Correlates with Its Functional Activities in Transcriptional Regulation and Infectivity in Cultured Fibroblast Cells. J. Virol. 78: 6527-6542 [Abstract] [Full Text]  
  • Terauchi, M., Koi, H., Hayano, C., Toyama-Sorimachi, N., Karasuyama, H., Yamanashi, Y., Aso, T., Shirakata, M. (2003). Placental Extravillous Cytotrophoblasts Persistently Express Class I Major Histocompatibility Complex Molecules after Human Cytomegalovirus Infection. J. Virol. 77: 8187-8195 [Abstract] [Full Text]  
  • Everett, R. D., Sourvinos, G., Orr, A. (2003). Recruitment of Herpes Simplex Virus Type 1 Transcriptional Regulatory Protein ICP4 into Foci Juxtaposed to ND10 in Live, Infected Cells. J. Virol. 77: 3680-3689 [Abstract] [Full Text]  
  • Stanton, R., Fox, J. D., Caswell, R., Sherratt, E., Wilkinson, G. W. G. (2002). Analysis of the human herpesvirus-6 immediate-early 1 protein. J. Gen. Virol. 83: 2811-2820 [Abstract] [Full Text]  
  • Noris, E., Zannetti, C., Demurtas, A., Sinclair, J., De Andrea, M., Gariglio, M., Landolfo, S. (2002). Cell Cycle Arrest by Human Cytomegalovirus 86-kDa IE2 Protein Resembles Premature Senescence. J. Virol. 76: 12135-12148 [Abstract] [Full Text]  
  • Xu, Y., Colletti, K. S., Pari, G. S. (2002). Human Cytomegalovirus UL84 Localizes to the Cell Nucleus via a Nuclear Localization Signal and Is a Component of Viral Replication Compartments. J. Virol. 76: 8931-8938 [Abstract] [Full Text]  
  • Hofmann, H., Sindre, H., Stamminger, T. (2002). Functional Interaction between the pp71 Protein of Human Cytomegalovirus and the PML-Interacting Protein Human Daxx. J. Virol. 76: 5769-5783 [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]  
  • Shirakata, M., Terauchi, M., Ablikim, M., Imadome, K.-I., Hirai, K., Aso, T., Yamanashi, Y. (2002). Novel Immediate-Early Protein IE19 of Human Cytomegalovirus Activates the Origin Recognition Complex I Promoter in a Cooperative Manner with IE72. J. Virol. 76: 3158-3167 [Abstract] [Full Text]  
  • Xu, Y., Ahn, J.-H., Cheng, M., apRhys, C. M., Chiou, C.-J., Zong, J., Matunis, M. J., Hayward, G. S. (2001). Proteasome-Independent Disruption of PML Oncogenic Domains (PODs), but Not Covalent Modification by SUMO-1, Is Required for Human Cytomegalovirus Immediate-Early Protein IE1 To Inhibit PML-Mediated Transcriptional Repression. J. Virol. 75: 10683-10695 [Abstract] [Full Text]  
  • Liao, G., Wu, F. Y., Hayward, S. D. (2001). Interaction with the Epstein-Barr Virus Helicase Targets Zta to DNA Replication Compartments. J. Virol. 75: 8792-8802 [Abstract] [Full Text]  
  • Bell, P., Montaner, L. J., Maul, G. G. (2001). Accumulation and Intranuclear Distribution of Unintegrated Human Immunodeficiency Virus Type 1 DNA. J. Virol. 75: 7683-7691 [Abstract] [Full Text]  
  • Ahn, J.-H., Xu, Y., Jang, W.-J., Matunis, M. J., Hayward, G. S. (2001). Evaluation of Interactions of Human Cytomegalovirus Immediate-Early IE2 Regulatory Protein with Small Ubiquitin-Like Modifiers and Their Conjugation Enzyme Ubc9. J. Virol. 75: 3859-3872 [Abstract] [Full Text]  
  • Adamson, A. L., Kenney, S. (2001). Epstein-Barr Virus Immediate-Early Protein BZLF1 Is SUMO-1 Modified and Disrupts Promyelocytic Leukemia Bodies. J. Virol. 75: 2388-2399 [Abstract] [Full Text]  
  • Meier, J. L. (2001). Reactivation of the Human Cytomegalovirus Major Immediate-Early Regulatory Region and Viral Replication in Embryonal NTera2 Cells: Role of Trichostatin A, Retinoic Acid, and Deletion of the 21-Base-Pair Repeats and Modulator. J. Virol. 75: 1581-1593 [Abstract] [Full Text]  
  • Wu, F. Y., Ahn, J.-H., Alcendor, D. J., Jang, W.-J., Xiao, J., Hayward, S. D., Hayward, G. S. (2001). Origin-Independent Assembly of Kaposi's Sarcoma-Associated Herpesvirus DNA Replication Compartments in Transient Cotransfection Assays and Association with the ORF-K8 Protein and Cellular PML. J. Virol. 75: 1487-1506 [Abstract] [Full Text]  
  • Bell, P., Lieberman, P. M., Maul, G. G. (2000). Lytic but Not Latent Replication of Epstein-Barr Virus Is Associated with PML and Induces Sequential Release of Nuclear Domain 10 Proteins. J. Virol. 74: 11800-11810 [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»