A nuclear DNA-binding protein expressed during early stages of B cell differentiation interacts with diverse segments within and 3' of the Ig H chain gene cluster

F. Liao, S. L. Giannini, B. K. Birshtein

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

We have used electrophoretic mobility shift assays (EMSA) to detect B cell lineage-specific nuclear proteins that bind to diverse segments within and 3' of the Ig H chain gene cluster. DNA binding sites include sequences 5' of each of the following C region genes: μ, γ1, γ2a, ε, and α. For the most part, these binding sites lie 5' of C(H)-associated tandem repeats. Binding sites for the same B cell lineage-specific proteins have also been defined in the region 3' of Cα, close to a recently described B cell-specific enhancer element. Cross-competition of EMSA indicates that the B cell lineage-specific nucleoprotein is indistinguishable from those described previously by others: Sα-BP and BSAP. Because of the diverse sequences recognized by this protein, we term it NF-HB, B-lineage-specific nuclear factor that binds to IgH gene segments. EMSA using segments 5' of Sγ2a (5'Sγ2a-176) and 3' of Cα (3'α- 88) shows multiple binding complexes, two of which are B cell lineage specific. The B cell-specific complex with fastest mobility contains only NF- HB, and the one with slowest mobility contains NF-HB together with a ubiquitous DNA-binding protein(s). The ubiquitous binding protein is different for 5'Sγ2a-176 and for 3'α-88, representing the formation of protein-NF-HB complexes specific for these particular Ig DNA regions. Spleen cells show a single band upon EMSA with either 5'Sγ2a-176 or 3'α-88. Upon LPS stimulation, additional binding complexes of slower mobility were formed resulting in a pattern comparable to those detected in pro-B, pre-B, and B cell lines. We hypothesize that NF-HB may promote physical interactions between the 3'α-enhancer and segments of the Ig H gene cluster.

Original languageEnglish (US)
Pages (from-to)2909-2917
Number of pages9
JournalJournal of Immunology
Volume148
Issue number9
StatePublished - 1992

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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