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. 1993 Jun;13(6):3611–3622. doi: 10.1128/mcb.13.6.3611

NF-HB (BSAP) is a repressor of the murine immunoglobulin heavy-chain 3' alpha enhancer at early stages of B-cell differentiation.

M Singh 1, B K Birshtein 1
PMCID: PMC359830  PMID: 8497273

Abstract

We have identified a nuclear factor expressed in pro-B-, pre-B-, and B-cell lines that binds to two sites within the murine immunoglobulin heavy-chain (IgH) 3' alpha enhancer (3' alpha E). These sites were defined by oligonucleotide competition in an electrophoretic mobility shift assay (EMSA) and methylation interference footprinting. The 3' alpha E-binding factor is indistinguishable from NF-HB (B-lineage-specific nuclear factor that binds to the IgH gene) and the B-lineage-specific transcription factor BSAP by several criteria, including similar cell type distribution of binding activity, cross-competition of binding sites in EMSA, similar protein size as demonstrated by UV cross-linking, and sequence identity of one of the 3' alpha E-binding sites with a BSAP-binding site within the promoter of the sea urchin late histone gene H2A-2.2. These observations indicate that 3' alpha E is one of the mammalian targets for NF-HB (BSAP). Transient-transfection assays with chloramphenicol acetyltransferase gene constructs containing 3' alpha E and mutant 3' alpha E, in which one of the NF-HB binding sites was inactivated by site-specific mutagenesis, showed ca. five- to sixfold-enhanced activity of mutated 3' alpha E over parental 3' alpha E in B-cell lines (NF-HB+), while no significant difference was observed in plasmacytoma cells (NF-HB-). We conclude from these observations that NF-HB (BSAP) acts as a repressor of the mouse IgH 3' alpha E.

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