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
Google Scholar
Right arrow Articles by Dallaire, F.
Right arrow Articles by Branton, P. E.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dallaire, F.
Right arrow Articles by Branton, P. E.

 Previous Article  |  Next Article 

Journal of Virology, December 2009, p. 12172-12184, Vol. 83, No. 23
0022-538X/09/$08.00+0     doi:10.1128/JVI.01169-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

A Proteomic Approach To Identify Candidate Substrates of Human Adenovirus E4orf6-E1B55K and Other Viral Cullin-Based E3 Ubiquitin Ligases {triangledown}

Frédéric Dallaire,1 Paola Blanchette,1 and Philip E. Branton1,2,3*

Departments of Biochemistry,1 Oncology,2 Goodman Cancer Center, McGill University, McIntyre Medical Building, 3655 Promenade Sir William Osler, Montreal, Quebec H3G 1Y6, Canada3

Received 8 June 2009/ Accepted 4 September 2009

It has been known for some time that the human adenovirus serotype 5 (Ad5) E4orf6 and E1B55K proteins work in concert to degrade p53 and to regulate selective export of late viral mRNAs during productive infection. Both of these functions rely on the formation by the Ad5 E4orf6 protein of a cullin 5-based E3 ubiquitin ligase complex containing elongins B and C. E1B55K is believed to function as the substrate recognition module for the complex and, in addition to p53, Mre11 and DNA ligase IV have also been identified as substrates. To discover additional substrates we have taken a proteomic approach by using two-dimensional difference gel electrophoresis to detect cellular proteins that decrease significantly in amount in p53-null H1299 human lung carcinoma cells after expression of E1B55K and E4orf6 using adenovirus vectors. Several species were detected and identified by mass spectroscopy, and for one of these, integrin {alpha}3, we went on in a parallel study to confirm it as a bone fide substrate of the complex (F. Dallaire et al., J. Virol. 83:5329-5338, 2009). Although the system has some limitations, it may still be of some general use in identifying candidate substrates of any viral cullin-based E3 ubiquitin ligase complex, and we suggest a series of criteria for substrate validation.

* Corresponding author. Mailing address: Department of Biochemistry, McGill University, McIntyre Medical Building, 3655 Promenade Sir William Osler, Montreal, Quebec H3G 1Y6, Canada. Phone: (514) 398-7268. Fax: (514) 398-7384. E-mail: philip.branton{at}mcgill.ca

{triangledown} Published ahead of print on 16 September 2009.

Journal of Virology, December 2009, p. 12172-12184, Vol. 83, No. 23
0022-538X/09/$08.00+0     doi:10.1128/JVI.01169-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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