Dyad Symmetry Within the Mouse 3′ IgH Regulatory Region Includes Two Virtually Identical Enhancers (Cα3′E and hs3)

The transcription of the murine Ig heavy chain locus is regulated not only by the intronic enhancer, Eμ, but also by a 3′ regulatory region located downstream of the Cα membrane exon. Several DNase I-hypersensitive sites (hs1-4) and enhancer elements (e.g., Cα3′E) have been identified in this 3′ regulatory region, and some of these were suggested to comprise a locus control region. However, little is known about the coordinate regulation or function of these individual elements. Here we provide evidence that Cα3′E and hs3 are virtually mirror images of each other and demarcate the edges of an ∼25-kb region of quasi-dyad symmetry with 3′αE(hs1,2) at its center. Flanking 3′αE(hs1,2) are inverted repeats and families of repetitive sequences uniquely located in this region. We have observed that, like 3′αE(hs1,2) and hs3, Cα3′E is DNase I hypersensitive in plasma cell lines, but not in a pre-B cell line. Additionally, we found that Cα3′E and hs3 show significant transcriptional synergy in transfection assays only in a plasma cell line. The DNA topology of the 3′ regulatory region coupled with new and existing data on the activity of its individual enhancers during B cell differentiation lead us to propose a biphasic model for the activity of this region. According to our model, one unit, consisting of the 3′-most enhancer, hs4, is active early and throughout B cell development. The second unit, which comprises Cα3′E, 3′αE(hs1,2), and hs3, becomes active later in development, when it contributes to such processes as class switching and increased levels of Ig heavy chain gene transcription in plasma cells.