Synergistic Activation of the Germline ε Promoter Mediated by Stat6 and C/EBPβ

Abstract

Transcription of the Ig H chain germline transcripts is a prerequisite for class switching. Expression of the ε germline transcript is induced by IL-4 and requires the integrity of a composite IL-4 response element. The element is bound by the IL-4-inducible transcription factor Stat6 and one or more members of the CAAT/enhancer-binding protein (C/EBP) family, a constitutively expressed class of transcription factors. Here, we show that Stat6 and C/EBPβ cooperate to synergistically activate transcription from the ε element. The effect was most pronounced in lymphoid cells, and the activation domains of both proteins were required to achieve this synergy. Although other members of the C/EBP family are able to bind the element, very little cooperativity was seen with C/EBPα and none with C/EBPγ. In fact, C/EBPγ was able to inhibit IL-4-induced reporter activity. Stat6 and C/EBPβ bind the IL-4 response element simultaneously. The fast dissociation rate apparent when Stat6 binds this DNA element alone is slowed when C/EBPβ binds at the neighboring site. These data suggest a mechanism whereby C/EBPβ stabilizes Stat6 binding at this element, thereby increasing the likelihood that both of their activation domains will interact, possibly with other factors, to activate transcription in an IL-4-dependent manner.