separation of anti-proliferation and anti-apoptotic functions of retinoblastoma protein through targeted mutations of its ab domain防扩散分离和抗凋亡功能的视网膜母细胞瘤蛋白通过有针对性的突变的ab域.pdfVIP

Separation of Anti-Proliferation and Anti-Apoptotic Functions of Retinoblastoma Protein through Targeted Mutations of Its A/B Domain B. Nelson Chau, Chris W. Pan, Jean Y. J. Wang* Department of Medicine, Division of Hematology/Oncology, Moores-UCSD Cancer Center, University of California San Diego, La Jolla, California, United States of America 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 (KN) mutant to disrupt the E2F- and LxCxE-binding pockets. The RB-K530A (K) mutant is sufficient to inactivate the E2F-binding pocket, whereas the RB-N757F (N) 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. Citation: Chau BN, Pan CW, Wang JYJ (2006) Separation of Anti-Proliferation and Anti-Apoptotic Functions of Retinoblastoma Protein through Targeted Mutations of Its A/B Domain. PLoS ONE 1(1): e82. doi:10.1371/journal.pone.0000082 INTRODUCTION