EVEN-SKIPPED HOMEOBOX 1 controls human ES cell differentiation by directly repressing GOOSECOID expression

TGFß signaling patterns the primitive streak, yet little is known about transcriptional effectors that mediate the cell fate choices during streak-like development in mammalian embryos and in embryonic stem (ES) cells. Here we demonstrate that cross-antagonistic actions of EVEN-SKIPPED HOMEOBOX 1 (EVX1) and GOOSECOID (GSC) regulate cell fate decisions in streak-like progenitors derived from human ES cells exposed to BMP4 and/or activin. We found that EVX1 repressed GSC expression and promoted formation of posterior streak-like progeny in response to BMP4, and conversely that GSC repressed EVX1 expression and was required for development of anterior streak-like progeny in response to activin. Chromatin immunoprecipitation assays showed that EVX1 bound to the GSC 5′-flanking region in BMP4 treated human ES cells, and band shift assays identified two EVX1 binding sites in the GSC 5′-region. Significantly, we found that intact EVX1 binding sites were required for BMP4-mediated repression of GSC reporter constructs. We conclude that BMP4-induced EVX1 repress GSC directly and the two genes form the core of a gene regulatory network (GRN) controlling cell fates in streak-like human ES cell progeny.

► EVX1 repress GSC and promotes formation of posterior streak-like cells in response to BMP4.
► GSC repress EVX1 and promotes formation of anterior streak-like cells in response to activin.
► EVX1 repress GSC directly via two functional EVX1 binding sites in the GSC 5′-region.
► EVX1 and GSC forms the core of a GRN controlling cell fates in streak-like ES cell progeny.