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 Boivin, D.
Right arrow Articles by Branton, P. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Boivin, D.
Right arrow Articles by Branton, P. E.

 Previous Article  |  Next Article 

Journal of Virology, February 1999, p. 1245-1253, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Analysis of Synthesis, Stability, Phosphorylation, and Interacting Polypeptides of the 34-Kilodalton Product of Open Reading Frame 6 of the Early Region 4 Protein of Human Adenovirus Type 5

Dominique Boivin,1,dagger Megan R. Morrison,1 Richard C. Marcellus,1,dagger Emmanuelle Querido,1 and Philip E. Branton1,2,*

Departments of Biochemistry1 and Oncology,2 McGill University, Montréal, Québec, Canada H3G 1Y6

Received 9 March 1998/Accepted 27 October 1998

The 34-kDa early-region 4 open reading frame 6 (E4orf6) product of human adenovirus type 5 forms complexes with both the cellular tumor suppressor p53 and the viral E1B 55-kDa protein (E1B-55kDa). E4orf6 can inhibit p53 transactivation activity, as can E1B-55kDa, and in combination these viral proteins cause the rapid turnover of p53. In addition, E4orf6-55kDa complexes play a critical role at later times in the regulation of viral mRNA transport and shutoff of host cell protein synthesis. In the present study, we have further characterized some of the biological properties of E4orf6. Analysis of extracts from infected cells by Western blotting indicated that E4orf6, like E1A and E1B products, is present at high levels until very late times, suggesting that it is available to act throughout the infectious cycle. This pattern is similar to that of E4orf4 but differs markedly from that of another E4 product, E4orf6/7, which is present only transiently. Synthesis of E4orf6 is maximal at early stages but ceases completely with the onset of shutoff of host protein synthesis; however, it was found that unlike E4orf6/7, E4orf6 is very stable, thus allowing high levels to be maintained even at late times. E4orf6 was shown to be phosphorylated at low levels. Coimmunoprecipitation studies in cells lacking p53 indicated that E4orf6 interacts with a number of other proteins. Five of these were shown to be viral or virally induced proteins ranging in size from 102 to 27 kDa, including E1B-55kDa. One such species, of 72 kDa, was shown not to represent the E2 DNA-binding protein and thus remains to be identified. Another appeared to be the L4 100-kDa nonstructural adenovirus late product, but it appeared to be present nonspecifically and not as part of an E4orf6 complex. Apart from p53, three additional cellular proteins, of 84, 19, and 14 kDa were detected by using an adenovirus vector that expresses only E4orf6. The 19-kDa species and a 16-kDa cellular protein were also shown to interact with E4orf6/7. It is possible that complex formation with these viral and cellular proteins plays a role in one or more of the biological activities associated with E4orf6 and E4orf6/7.

* Corresponding author. Mailing address: Department of Biochemistry, McGill University, McIntyre Medical Sciences Building, 3655 Drummond St., Montréal, Québec, Canada H3G 1Y6. Phone: (514) 398-8350. Fax: (514) 398-7384. E-mail: branton{at}medcor.mcgill.ca

dagger Present address: Geminx Biotechnologies, Montréal, Québec, Canada H2W 2M9.

Journal of Virology, February 1999, p. 1245-1253, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Dallaire, F., Blanchette, P., Branton, P. E. (2009). A Proteomic Approach To Identify Candidate Substrates of Human Adenovirus E4orf6-E1B55K and Other Viral Cullin-Based E3 Ubiquitin Ligases. J. Virol. 83: 12172-12184 [Abstract] [Full Text]  
  • Kindsmuller, K., Schreiner, S., Leinenkugel, F., Groitl, P., Kremmer, E., Dobner, T. (2009). A 49-Kilodalton Isoform of the Adenovirus Type 5 Early Region 1B 55-Kilodalton Protein Is Sufficient To Support Virus Replication. J. Virol. 83: 9045-9056 [Abstract] [Full Text]  
  • Dallaire, F., Blanchette, P., Groitl, P., Dobner, T., Branton, P. E. (2009). Identification of Integrin {alpha}3 as a New Substrate of the Adenovirus E4orf6/E1B 55-Kilodalton E3 Ubiquitin Ligase Complex. J. Virol. 83: 5329-5338 [Abstract] [Full Text]  
  • Miron, M.-J., Blanchette, P., Groitl, P., Dallaire, F., Teodoro, J. G., Li, S., Dobner, T., Branton, P. E. (2009). Localization and Importance of the Adenovirus E4orf4 Protein during Lytic Infection. J. Virol. 83: 1689-1699 [Abstract] [Full Text]  
  • Schwartz, R. A., Lakdawala, S. S., Eshleman, H. D., Russell, M. R., Carson, C. T., Weitzman, M. D. (2008). Distinct Requirements of Adenovirus E1b55K Protein for Degradation of Cellular Substrates. J. Virol. 82: 9043-9055 [Abstract] [Full Text]  
  • Blanchette, P., Kindsmuller, K., Groitl, P., Dallaire, F., Speiseder, T., Branton, P. E., Dobner, T. (2008). Control of mRNA Export by Adenovirus E4orf6 and E1B55K Proteins during Productive Infection Requires E4orf6 Ubiquitin Ligase Activity. J. Virol. 82: 2642-2651 [Abstract] [Full Text]  
  • Kindsmuller, K., Groitl, P., Hartl, B., Blanchette, P., Hauber, J., Dobner, T. (2007). Intranuclear targeting and nuclear export of the adenovirus E1B-55K protein are regulated by SUMO1 conjugation. Proc. Natl. Acad. Sci. USA 104: 6684-6689 [Abstract] [Full Text]  
  • Gonzalez, R., Huang, W., Finnen, R., Bragg, C., Flint, S. J. (2006). Adenovirus E1B 55-Kilodalton Protein Is Required for both Regulation of mRNA Export and Efficient Entry into the Late Phase of Infection in Normal Human Fibroblasts. J. Virol. 80: 964-974 [Abstract] [Full Text]  
  • Blanchette, P., Cheng, C. Y., Yan, Q., Ketner, G., Ornelles, D. A., Dobner, T., Conaway, R. C., Conaway, J. W., Branton, P. E. (2004). Both BC-Box Motifs of Adenovirus Protein E4orf6 Are Required To Efficiently Assemble an E3 Ligase Complex That Degrades p53. Mol. Cell. Biol. 24: 9619-9629 [Abstract] [Full Text]  
  • Mohammadi, E. S., Ketner, E. A., Johns, D. C., Ketner, G. (2004). Expression of the adenovirus E4 34k oncoprotein inhibits repair of double strand breaks in the cellular genome of a 293-based inducible cell line. Nucleic Acids Res 32: 2652-2659 [Abstract] [Full Text]  
  • Harada, J. N., Shevchenko, A., Shevchenko, A., Pallas, D. C., Berk, A. J. (2002). Analysis of the Adenovirus E1B-55K-Anchored Proteome Reveals Its Link to Ubiquitination Machinery. J. Virol. 76: 9194-9206 [Abstract] [Full Text]  
  • Gonzalez, R. A., Flint, S. J. (2002). Effects of Mutations in the Adenoviral E1B 55-Kilodalton Protein Coding Sequence on Viral Late mRNA Metabolism. J. Virol. 76: 4507-4519 [Abstract] [Full Text]  
  • Orlando, J. S., Ornelles, D. A. (2002). E4orf6 Variants with Separate Abilities To Augment Adenovirus Replication and Direct Nuclear Localization of the E1B 55-Kilodalton Protein. J. Virol. 76: 1475-1487 [Abstract] [Full Text]  
  • Querido, E., Blanchette, P., Yan, Q., Kamura, T., Morrison, M., Boivin, D., Kaelin, W. G., Conaway, R. C., Conaway, J. W., Branton, P. E. (2001). Degradation of p53 by adenovirus E4orf6 and E1B55K proteins occurs via a novel mechanism involving a Cullin-containing complex. Genes Dev. 15: 3104-3117 [Abstract] [Full Text]  
  • Querido, E., Morisson, M. R., Chu-Pham-Dang, H., Thirlwell, S. W.-L., Boivin, D., Branton, P. E. (2001). Identification of Three Functions of the Adenovirus E4orf6 Protein That Mediate p53 Degradation by the E4orf6-E1B55K Complex. J. Virol. 75: 699-709 [Abstract] [Full Text]  
  • Cathomen, T., Weitzman, M. D. (2000). A Functional Complex of Adenovirus Proteins E1B-55kDa and E4orf6 Is Necessary To Modulate the Expression Level of p53 but Not Its Transcriptional Activity. J. Virol. 74: 11407-11412 [Abstract] [Full Text]  
  • Russell, W. C. (2000). Update on adenovirus and its vectors. J. Gen. Virol. 81: 2573-2604 [Full Text]  
  • Nevels, M., Rubenwolf, S., Spruss, T., Wolf, H., Dobner, T. (2000). Two Distinct Activities Contribute to the Oncogenic Potential of the Adenovirus Type 5 E4orf6 Protein. J. Virol. 74: 5168-5181 [Abstract] [Full Text]  
  • Grifman, M., Chen, N. N., Gao, G.-p., Cathomen, T., Wilson, J. M., Weitzman, M. D. (1999). Overexpression of Cyclin A Inhibits Augmentation of Recombinant Adeno-Associated Virus Transduction by the Adenovirus E4orf6 Protein. J. Virol. 73: 10010-10019 [Abstract] [Full Text]  
  • Lusky, M., Grave, L., Dieterle, A., Dreyer, D., Christ, M., Ziller, C., Furstenberger, P., Kintz, J., Ali Hadji, D., Pavirani, A., Mehtali, M. (1999). Regulation of Adenovirus-Mediated Transgene Expression by the Viral E4 Gene Products: Requirement for E4 ORF3. J. Virol. 73: 8308-8319 [Abstract] [Full Text]  
  • Boyer, J. L., Ketner, G. (2000). Genetic Analysis of a Potential Zinc-binding Domain of the Adenovirus E4 34k Protein. J. Biol. Chem. 275: 14969-14978 [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»