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J Virol, March 1998, p. 2323-2334, Vol. 72, No. 3
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Anchorage-Independent Transcription of the Cyclin A Gene Induced by the E7 Oncoprotein of Human Papillomavirus Type 16

Almut Schulze, Boris Mannhardt, Karin Zerfass-Thome, Werner Zwerschke, and Pidder Jansen-Dürr*

Forschungsschwerpunkt Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, D-69120 Heidelberg, Germany

Received 24 July 1997/Accepted 12 December 1997

To develop an experimental model for E7-mediated anchorage-independent growth, we studied the ability of E7-expressing NIH 3T3 subclones to enter S phase when they were cultured in suspension. We found that expression of E7 prevents the inhibition of cyclin E-associated kinase and also triggers activation of cyclin A gene expression in suspension cells. A point mutation in the amino terminus of E7 prevented E7-driven rescue of cyclin E-associated kinase activity in suspension cells; however, cells with this mutation retained some ability to activate cyclin A gene expression and promote S-phase entry. Activation of cyclin A gene expression by E7 was correlated with an increased binding of free E2F to a regulatory element in the cyclin A promoter which mediates both repression of cyclin A upon loss of adhesion and its reactivation by E7. Surprisingly, expression of E7 led to a nuclear accumulation of one species of free E2F, namely, an E2F-4-DP-1 heterodimer, that is exclusively cytoplasmic in the absence of E7. Taken together, the data reported here indicate that several different E7-dependent changes of cellular-growth-regulating pathways can cooperate to allow adhesion-independent entry into S phase.

* Corresponding author. Mailing address: Forschungsschwerpunkt Angewandt Tumorvirologie, Abt. 620, INF 242, D-69120 Heidelberg, Germany. Phone: 49-6221-424628. Fax: 49-6221-424902. E-mail: p.jansen-duerr{at}dkfz-heidelberg.de.




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