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3 (AML2) Is Induced by TGF-ß1 to Bind and Activate the Mouse Germline Ig Promoter1
Department of Molecular Genetics and Microbiology, Graduate Program in Immunology and Virology, University of Massachusetts Medical School, Worcester, MA 01655
TGF-ß1 directs class switching to IgA by splenic B cells and by
the surface IgM+ B cell line, I.29µ, by inducing germline
(GL) Ig 
transcripts. The promoter segment between -130 and +46,
relative to the first initiation site for mouse GL transcripts, is
sufficient for expression and TGF-ß1 inducibility of a reporter gene
in B cell lines. Within this segment resides a TGF-ß1-responsive
element (TßRE) that is required for induction of the promoter by
TGF-ß1 and, when multimerized, is sufficient to transfer TGF-ß1
inducibility to another promoter. In this report we show that a
TGF-ß1-inducible complex binds the TßRE and contains the
transcription factor core-binding factor (CBF; also known as acute
myeloid leukemia, AML). Although all three CBF family members
activate the GL promoter, only CBF3 (AML-2) is induced by
TGF-ß1 in splenic B and I.29µ cells. The TßRE contains two CBF
binding sites. Mutation of both sites reduces but does not eliminate
induction of the GL promoter by TGF-ß1 or by overexpression of
CBF, possibly due to the presence of an additional CBF site in the
promoter. In addition, the TßRE contains two copies of another
sequence motif. Mutation of these motifs eliminates TGF-ß1 induction
of the GL promoter. Together the data indicate that TGF-ß1
induction of the promoter involves induction of CBF3, which
binds to the TßRE of the promoter along with one or more
proteins.
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