IDENTIFICATION OF DISTINCT ROLES FOR SEPARATE E1A DOMAINS IN DISRUPTION OF E2F COMPLEXES

The adenovirus EIA protein can disrupt protein complexes containing the E2F transcription factor in association with cellular regulatory proteins such as the retinoblastoma gene product (Rb) and the Rb-related p107 protein. Previous experiments have shown that the CR1 and CR2 domains of E1A are required for this activity. We now demonstrate that the CR2 domain is essential for allowing E1A to interact with the E2F-Rb or the E2F-p107-cyclin A-cdk2 complex. Multimeric complexes containing E1A can be detected when the CRI domain has been rendered inactive by mutation. In addition, the EIA CR1 domain, but not the CR2 domain, is sufficient to prevent the interaction of E2F with Rb or p107. On the basis of these results, we suggest a model whereby the CR2 domain brings EIA to the E2F complexes and then, upon a normal equilibrium dissociation of Rb or p107 from E2F, the ElA CRI domain is able to block the site of interaction on Rb or p107, thereby preventing the re-formation of the complexes.