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

Fang Liao, Sandra L. Giannini, Barbara K. Birshtein

Research output: Contribution to journalArticle

48 Citations (Scopus)

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 CH-associated tandem repeats. Binding sites for the same B cell lineagespecific 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: Sa-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 Ig H gene segments. EMSA using segments 5′ of Sγ2a (5′Sγ2a-176) and 3′ of Ca (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, preB, 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 - May 1 1992

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DNA-Binding Proteins
Multigene Family
Nuclear Proteins
Cell Differentiation
B-Lymphocytes
Electrophoretic Mobility Shift Assay
Cell Lineage
Immunoglobulin Genes
Binding Sites
Proteins
Tandem Repeat Sequences
Nucleoproteins
DNA
Carrier Proteins
Spleen
Cell Line
Genes

ASJC Scopus subject areas

  • Immunology

Cite this

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. / Liao, Fang; Giannini, Sandra L.; Birshtein, Barbara K.

In: Journal of Immunology, Vol. 148, No. 9, 01.05.1992, p. 2909-2917.

Research output: Contribution to journalArticle

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