Glycogen Synthase Kinase 3 Phosphorylates RBL2/p130 during Quiescence

Abstract

Phosphorylation of the retinoblastoma-related or pocket proteins RB1/pRb, RBL1/p107, and RBL2/p130 regulates cell cycle progression and exit. While all pocket proteins are phosphorylated by cyclin-dependent kinases (CDKs) during the G₁/S-phase transition, p130 is also specifically phosphorylated in G₀-arrested cells. We have previously identified several phosphorylated residues that match the consensus site for glycogen synthase kinase 3 (GSK3) in the G₀ form of p130. Using small-molecule inhibitors of GSK3, site-specific mutants of p130, and phospho-specific antibodies, we demonstrate here that GSK3 phosphorylates p130 during G₀. Phosphorylation of p130 by GSK3 contributes to the stability of p130 but does not affect its ability to interact with E2F4 or cyclins. Regulation of p130 by GSK3 provides a novel link between growth factor signaling and regulation of the cell cycle progression and exit.

We are grateful to William Hahn, Ralph Scully, and Hartmut Land for reagents.

This work was supported in part by Public Health Service grant RO1-CA63113 (J.D.).