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Published online on April 18, 2000, 10.1073/pnas.080078197
PNAS | April 25, 2000 | vol. 97 | no. 9 | 4879-4884


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Vol. 97, Issue 9, 4879-4884, April 25, 2000

Microbiology
 Genome-wide analysis of vaccinia virus protein-protein interactions

Stephen McCraith*, Ted Holtzmandagger , Bernard MossDagger , and Stanley Fields*,§,

* Departments of Genetics and Medicine, Box 357360, dagger  Locke Computer Center, and § Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195; and Dagger  Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0445

Contributed by Bernard Moss, February 23, 2000

To detect interactions between proteins of vaccinia virus, we carried out a comprehensive two-hybrid analysis to assay every pairwise combination. We constructed an array of yeast transformants that contained each of the 266 predicted viral ORFs as Gal4 activation domain hybrid proteins. The array was individually mated to transformants containing each ORF as a Gal4-DNA-binding domain hybrid, and diploids expressing the two-hybrid reporter gene were identified. Of the approx 70,000 combinations, we found 37 protein-protein interactions, including 28 that were previously unknown. In some cases, e.g., late transcription factors, both proteins were known to have related roles although there was no prior evidence of physical associations. For some other interactions, neither protein had a known role. In the majority of cases, however, one of the interacting proteins was known to be involved in DNA replication, transcription, virion structure, or host evasion, thereby providing a clue to the role of the other uncharacterized protein in a specific process.

To whom reprint requests should be addressed at: Howard Hughes Medical Institute, Departments of Genetics and Medicine, University of Washington, Box 357360, Seattle, WA 98195. E-mail: fields{at}u.washington.edu.


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