Interchangeable Roles for E2F Transcriptional Repression by the Retinoblastoma Protein and p27KIP1-Cyclin-Dependent Kinase Regulation in Cell Cycle Control and Tumor Suppression

Abstract

The mammalian G₁-S phase transition is controlled by the opposing forces of cyclin-dependent kinases (CDK) and the retinoblastoma protein (pRB). Here, we present evidence for systems-level control of cell cycle arrest by pRB-E2F and p27-CDK regulation. By introducing a point mutant allele of pRB that is defective for E2F repression (Rb1^G) into a p27^KIP1 null background (Cdkn1b^-/-), both E2F transcriptional repression and CDK regulation are compromised. These double-mutant Rb1^G/G; Cdkn1b^-/- mice are viable and phenocopy Rb1^+/- mice in developing pituitary adenocarcinomas, even though neither single mutant strain is cancer prone. Combined loss of pRB-E2F transcriptional regulation and p27^KIP1 leads to defective proliferative control in response to various types of DNA damage. In addition, Rb1^G/G; Cdkn1b^-/- fibroblasts immortalize faster in culture and more frequently than either single mutant genotype. Importantly, the synthetic DNA damage arrest defect caused by Rb1^G/G; Cdkn1b^-/- mutations is evident in the developing intermediate pituitary lobe where tumors ultimately arise. Our work identifies a unique relationship between pRB-E2F and p27-CDK control and offers in vivo evidence that pRB is capable of cell cycle control through E2F-independent effects.

Keywords: CDK; DNA damage; DNA damage checkpoints; E2F; cell cycle; cyclin-dependent kinases; tumor suppressor; tumor suppressor genes.