An E2F Binding-Deficient Rb1 Protein Partially Rescues Developmental Defects Associated with Rb1 Nullizygosity

Abstract

Rb1 is essential for normal embryonic development, as null mice die in midgestation with widespread unscheduled cell proliferation. Rb1 protein (pRb) mediates cell cycle control by binding E2F transcription factors and repressing expression from E2F-dependent promoters. An increasing amount of evidence suggests that pRb loss also compromises cellular differentiation. Since differentiation is often dependent on cell cycle exit, it is currently unclear whether the effects of pRb on differentiation are an indirect consequence of pRb/E2F-mediated cell cycle control or whether they reflect direct cell-type-specific pRb functions. We have mutated Rb1 in the mouse to express a protein (R654W) specifically deficient in binding E2F1, E2F2, and E2F3. R654W mutant embryos exhibit cell cycle defects the same as those of Rb1 null embryos, reinforcing the importance of the interactions of pRb with E2F1, E2F2, and E2F3 for cell cycle control. However, R654W embryos survive at least 2 days longer than Rb1 null embryos, and increased life span is associated with improved erythrocyte and fetal liver macrophage differentiation. In contrast, R654W pRb does not rescue differentiation defects associated with pRb-deficient retinae. These data indicate that Rb1 makes important cell-type-specific contributions to cellular differentiation that are genetically separable from its general ability to stably bind E2F1, E2F2, and E2F3 and regulate the cell cycle.

Supplemental material for this article may be found at http://mcb.asm.org/.

We thank Paul Soloway (Cornell University) and John Clifford (MD Anderson Cancer Center) for sharing plasmids. We acknowledge Karin Williams, Harold Love, Mary Vaughan, Jiakun Zhang, and Leyfou Dabo for technical advice. We acknowledge the Vanderbilt Mouse Metabolic Physiology Center for performing the fatty acid analysis.

This work was supported by grants from the NIH to D.W.G. (CA70292), S.W.H. (CA96403), and M.A.D. (EY014867). The Charlotte Geyer Foundation (D.W.G.), the American Lebanese Syrian Associated Charities (M.A.D.), and Research to Prevent Blindness (M.A.D.) also supported the work described. M.A.D. is a Pew Scholar.