ETS1 is a genome-wide effector of RAS/ERK signaling in epithelial cells

Abstract

The RAS/ERK pathway is commonly activated in carcinomas and promotes oncogenesis by altering transcriptional programs. However, the array of cis-regulatory elements and trans-acting factors that mediate these transcriptional changes is still unclear. Our genome-wide analysis determined that a sequence consisting of neighboring ETS and AP-1 transcription factor binding sites is enriched near cell migration genes activated by RAS/ERK signaling in epithelial cells. In vivo screening of candidate ETS proteins revealed that ETS1 is specifically required for migration of RAS/ERK activated cells. Furthermore, both migration and transcriptional activation through ETS/AP-1 required ERK phosphorylation of ETS1. Genome-wide mapping of multiple ETS proteins demonstrated that ETS1 binds specifically to enhancer ETS/AP-1 sequences. ETS1 occupancy, and its role in cell migration, was conserved in epithelial cells derived from multiple tissues, consistent with a chromatin organization common to epithelial cell lines. Genome-wide expression analysis showed that ETS1 was required for activation of RAS-regulated cell migration genes, but also identified a surprising role for ETS1 in the repression of genes such as DUSP4, DUSP6 and SPRY4 that provide negative feedback to the RAS/ERK pathway. Consistently, ETS1 was required for robust RAS/ERK pathway activation. Therefore, ETS1 has dual roles in mediating epithelial-specific RAS/ERK transcriptional functions.

SLFN11 Is a Transcriptional Target of EWS-FLI1 and a Determinant of Drug Response in Ewing Sarcoma

“…ETS1 has been shown to play an important role in the progression of breast cancer (46) and has been recently linked to the RAS/ERK pathway in carcinomas (47). Bonetti et al recently demonstrated that FLI1 and ETS1 cooperatively contribute to the growth of diffuse large B-cell lymphoma, and influence gene expression related to germinal center differentiation (48).…”

An Interaction with Ewing’s Sarcoma Breakpoint Protein EWS Defines a Specific Oncogenic Mechanism of ETS Factors Rearranged in Prostate Cancer

“…The GGAA repeat reporter was created by annealing complementary DNA oligonucleotides (See Supplemental Experimental Procedures) containing 7 tandem repeats of GGAA or GAGA and cloning into pGL4.25. Dual luciferase reporter assay (Promega) measured luciferase activity as described (Plotnik et al, 2014). Firefly luciferase values were normalized to renilla values.…”

“…These ETS proteins activated transcription of cell migration genes by binding cis-regulatory sequences that have neighboring ETS and AP-1 transcription factor binding sites. However, this ETS/AP-1 binding is not specific to ERG, ETV1, ETV4, and ETV5, because ETS1 can also bind ETS/AP-1 sequences and activate transcription of cell migration genes in KRAS-mutant cancer cells (Plotnik et al, 2014). Therefore the molecular mechanism behind the specific biological function of ERG, ETV1, ETV4, and ETV5 in prostate cells is unknown.…”

Genome-Wide Analysis of RAS/ERK Signaling Targets

“…Here, we present a method for identifying gene expression programs regulated by RAS/ERK signaling using RNA-sequencing ( RNA-seq ) and differential gene expression analysis [ 7 ]. Using a MEK inhibitor , we disrupt the constitutive activation of the RAS/ RAF/MEK/ERK pathway in the DU145 prostate cancer cell line and then isolate mRNA .…”

“…1 for an overview of the main protocol steps). Using this approach, we have identifi ed nearly 700 genes signifi cantly regulated by RAS/ERK signaling, enriched in genes involved in cell migration , invasion, and proliferation [ 7 ].…”