Abstract

Pioneer transcription factors are DNA-binding proteins that can bind to nucleosomes in closed chromatin regions, exposing enhancers and promoters of genes for transcription. The action of these factors underpin stem cell pluripotency, cell reprogramming and differentiation. ELF2 is an ETS family pioneer factor that has a strong preference for oriented binding on nucleosomes at a composite head-to-tail dimeric sequence motif with a 2 bp spacing between the GGAA core elements. In this study, we investigated the interaction between ELF2 and a nucleosome using single-particle cryo-electron microscopy (cryo-EM). The ELF2-nucleosome structure shows two ELF2 proteins bound to the nucleosome cooperatively at superhelical location +4. The recognition [a]4 helices of both ELF2 monomers dock into the major groove at the core GGAA motifs and unwrap approximately four helical turns of the DNA from the surface of the nucleosome. The unwrapping almost completely exposes one histone H2A:H2B dimer, which dissociates in a subset of particles to form an ELF2-bound hexasome. ChIP-seq combined with genome-wide motif mapping indicates that all unmethylated high-affinity ELF2 double motifs are occupied, indicating that ELF2 is able to use its binding energy to function as a pioneer factor both in vitro and in vivo. ELF2 binding is highly enriched downstream of transcription start sites (TSS) of highly expressed genes, with the motifs oriented in such a way that the TSS becomes accessible upon ELF2 binding. This mechanism is essential for cells, as mutation of ELF2 motifs result in decreased cell proliferation. Taken together, our results indicate that ELF2 can use its binding energy to open chromatin in an oriented fashion, facilitating nucleosome positioning and transcription initiation at transcription start sites.