RBF Binding to both Canonical E2F Targets and Noncanonical Targets Depends on Functional dE2F/dDP Complexes

• DOI: 10.1128/mcb.00536-12
• Publication Date: 2012-08-28T03:12:15Z
• Authors: Michael Korenjak; Endre Anderssen; Sridhar Ramaswamy; Johnathan R. Whet...; Nicholas J. Dyson
• Keywords: Chromatin immunoprecipitation; E2F; ChIP-sequencing

The retinoblastoma (RB) family of proteins regulate transcription. These lack intrinsic DNA-binding activity but are recruited to specific genomic locations through interactions with sequence-specific factors. best-known target RB protein (pRB) is the E2F transcription factor; however, many other chromatin-associated have been described that may allow members act at additional sites. To gain a perspective on scale E2F-dependent and E2F-independent functions, we generated genome-wide binding profiles RBF1 dE2F in Drosophila larvae. dE2F2 associate large number sites genes diverse biological functions. In contrast, dE2F1 was detected smaller set promoters, suggesting it overrides repression by RBF1/dE2F2 subset targets. Approximately 15% RBF1-bound regions lacked consensus E2F-binding motifs. test whether action these independent, examined dDP mutant larvae any functional dE2F/dDP heterodimers. As measured chromatin immunoprecipitation-microarray analysis (ChIP-chip), ChIP-quantitative PCR (qPCR), cell fractionation, stable association eliminated mutants. This requirement for seen classic E2F-regulated promoters canonical results suggest E2F/DP complexes essential all targeting RBF1.