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Identification of E2A target genes in B lymphocyte development by using a gene tagging-based chromatin immunoprecipitation system.

Publication ,  Journal Article
Greenbaum, S; Zhuang, Y
Published in: Proc Natl Acad Sci U S A
November 12, 2002

The transcription factors encoded by the E2A gene are known to be essential for B lymphocyte development, and ectopic expression or gene inactivation studies have revealed several potential lineage-specific E2A target genes. However, it remains unknown whether these target genes are directly regulated by E2A at the transcriptional level. We therefore generated mice carrying an affinity-tagged E2A knock-in allele to provide a system for the direct elucidation of E2A target genes based on E2A binding to target regulatory regions. Abelson-transformed pre-B cell lines derived from these mice were used in chromatin immunoprecipitation experiments to identify regulatory sequences bound by E2A in the context of an early B lymphocyte environment. Significant E2A binding was detected at the promoters and enhancers of several essential B-lineage genes, including the Igkappa intronic and 3' enhancers, lambda5 and VpreB surrogate light chain promoters, the EBF locus promoter region, and the mb-1 (Igalpha) promoter. Low levels of E2A binding were observed at several other lymphoid-restricted regulatory regions including the Ig heavy chain (IgH) intronic enhancer, the IgH 3' enhancers hs3b/hs4, the RAG-2 enhancer, and the 5' regions of the B29 and TdT loci. An E2A target gene, the predicted butyrophilin-like gene NG9 (BTL-II), was also identified by using a chromatin immunoprecipitation-based cloning strategy. In summary, our studies have provided evidence that E2A is directly involved in the transcriptional regulation of a number of early B-lineage genes.

Duke Scholars

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

November 12, 2002

Volume

99

Issue

23

Start / End Page

15030 / 15035

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Transcription Factors
  • Regulatory Sequences, Nucleic Acid
  • Recombinant Fusion Proteins
  • Promoter Regions, Genetic
  • Polymerase Chain Reaction
  • Mice, Knockout
  • Mice
  • Humans
  • DNA-Binding Proteins
 

Citation

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Greenbaum, S., & Zhuang, Y. (2002). Identification of E2A target genes in B lymphocyte development by using a gene tagging-based chromatin immunoprecipitation system. Proc Natl Acad Sci U S A, 99(23), 15030–15035. https://doi.org/10.1073/pnas.232299999
Greenbaum, Stephen, and Yuan Zhuang. “Identification of E2A target genes in B lymphocyte development by using a gene tagging-based chromatin immunoprecipitation system.Proc Natl Acad Sci U S A 99, no. 23 (November 12, 2002): 15030–35. https://doi.org/10.1073/pnas.232299999.
Greenbaum, Stephen, and Yuan Zhuang. “Identification of E2A target genes in B lymphocyte development by using a gene tagging-based chromatin immunoprecipitation system.Proc Natl Acad Sci U S A, vol. 99, no. 23, Nov. 2002, pp. 15030–35. Pubmed, doi:10.1073/pnas.232299999.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

November 12, 2002

Volume

99

Issue

23

Start / End Page

15030 / 15035

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Transcription Factors
  • Regulatory Sequences, Nucleic Acid
  • Recombinant Fusion Proteins
  • Promoter Regions, Genetic
  • Polymerase Chain Reaction
  • Mice, Knockout
  • Mice
  • Humans
  • DNA-Binding Proteins