E μ and 3′RR IgH enhancers show hierarchic unilateral dependence in mature B-cells

Enhancer and super-enhancers are master regulators of cell fate. While they act at long-distances on adjacent genes, it is unclear whether they also act on one another. The immunoglobulin heavy chain (IgH) locus is unique in carrying two super-enhancers at both ends of the constant gene cluster: the 5′Eμ super-enhancer promotes VDJ recombination during the earliest steps of B-cell ontogeny while the 3′ regulatory region (3′RR) is essential for late differentiation. Since they carry functional synergies in mature B-cells and physically interact during IgH locus DNA looping, we investigated if they were independent engines of locus remodelling or if their function was more intimately intermingled, their optimal activation then requiring physical contact with each other. Analysis of chromatin marks, enhancer RNA transcription and accessibility in Eμ- and 3′RR-deficient mice show, in mature activated B-cells, an unilateral dependence of this pair of enhancers: while the 3′RR acts in autonomy, Eμ in contrast likely falls under control of the 3′RR.