Joy Wang

Background. The human retinoblastoma susceptibility gene encodes a nuclear phosphoprotein RB, which is a negative regulator of cell proliferation. The growth suppression function of RB requires an evolutionarily conserved A/B domain that contains two distinct peptide-binding pockets. At the A/B interface is a binding site for the C-terminal trans-activation domain of E2F. Within the B-domain is a binding site for proteins containing the LxCxE peptide motif. Methodology/Principle Findings. Based on the crystal structure of the A/ B domain, we have constructed an RB-K530A/N757F mutant to disrupt the E2F- and LxCxE-binding pockets. The RB-K530A mutant is sufficient to inactivate the E2F-binding pocket, whereas the RB-N757F mutant is sufficient to inactivate the LxCxE-binding pocket. Each single mutant inhibits cell proliferation, but the RB-KN double mutant is defective in growth suppression. Nevertheless, the RB-KN mutant is capable of reducing etoposide-induced apoptosis. Conclusion/Significance. Previous studies have established that RB-dependent G1-arrest can confer resistance to DNA damage-induced apoptosis. Results from this study demonstrate that RB can also inhibit apoptosis independent of growth suppression. © 2006 Chau et al.