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Journal of Virology, January 2000, p. 812-816, Vol. 74, No. 2
Institute of Pathology, Case Western Reserve
University School of Medicine, Cleveland, Ohio 44106
Received 2 July 1999/Accepted 15 October 1999
Papillomavirus E6 oncoproteins transform mammalian cells through
interaction with cellular proteins. Bovine papillomavirus type 1 E6
(BE6) interacts with three previously described cellular targets: the
E6AP E3 ubiquitin ligase, the calcium-binding protein E6BP (also known
as ERC-55), and paxillin, which is a focal adhesion adapter protein.
BE6 interacts strongly with each of these proteins in vitro, binding to
similar peptide sequences found in E6AP, E6BP, and paxillin. To
determine which BE6 interactions are necessary for transformation
by BE6, we used a novel selection strategy for temperature-sensitive
BE6 mutants in yeast that could discriminate in their interaction
between E6AP, E6BP, and paxillin. All BE6 mutants that retained
transforming ability retained association with paxillin, while some
mutants that were transformation positive failed to interact with E6AP
or E6BP. This study demonstrates that oncogene mutants that are
temperature sensitive for transformation can be selected in yeast and
that the induction of anchorage-independent cell proliferation by BE6
does not require strong association of BE6 with either E6AP or E6BP. Of
particular interest is the identification of a BE6 mutant
that interacts strongly with the acidic charged leucine motifs of E6AP,
E6BP, and paxillin but is devoid of transformation activity, thereby
genetically identifying a second essential transformation function in
BE6 that is independent of interaction with acidic charged leucine motifs.
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Identification of a Second Transforming Function in Bovine
Papillomavirus Type 1 E6 and the Role of E6 Interactions with
Paxillin, E6BP, and E6AP
*
Corresponding author. Mailing address: Department of
Pathology, Case Western Reserve University, 10900 Euclid Ave.,
Cleveland, OH 44106. Phone: (216) 368-1679. Fax: (216) 368-1300. E-mail: sbv{at}pop.cwru.edu.
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