Abstract
Genetic studies have demonstrated that the basic helix-loop-helix protein E2A is an essential transcription factor in B lymphocyte lineage commitment and differentiation. However, the mechanism underlying E2A-mediated transcription regulation is not fully understood. Here, we investigated the physical and genetic interactions between E2A and co-activators histone acetyltransferases (HATs) in B cells. Gel filtration analysis of human pre-B cell nuclear extract showed that E2A co-elutes with the HATs p300, CBP, and PCAF. A co-immunoprecipitation assay further demonstrated that a fraction of endogenous E2A proteins is associated with each of the three HATs. We show that these HATs acetylate E2A in vitro, enhance E2A-mediated transcription activity, and promote nuclear retention of E2A proteins. A catalytic mutation of p300 completely abrogates the ability of p300 to acetylate E2A and to promote E2A nuclear retention in 293T cells. A breeding test between E2A heterozygous mice and p300 heterozygous mice demonstrated that these two genes interact for proper B cell development. Collectively, these results suggest that E2A and HATs collaboratively regulate B cell development.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Acetyltransferases / metabolism*
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Age Factors
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Animals
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Animals, Newborn
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B-Lymphocytes / metabolism*
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B-Lymphocytes / physiology
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Basic Helix-Loop-Helix Transcription Factors
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Blotting, Western
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Cell Nucleus / metabolism
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Cell Separation
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Cells, Cultured
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Chromatography, Gel
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DNA-Binding Proteins / biosynthesis*
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DNA-Binding Proteins / genetics*
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E1A-Associated p300 Protein
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Flow Cytometry
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Gene Expression Regulation, Developmental*
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Genes, Reporter
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HeLa Cells
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Heterozygote
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Histone Acetyltransferases
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Humans
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Luciferases / metabolism
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Mice
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Microscopy, Fluorescence
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Mutation
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Nuclear Proteins / metabolism
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Precipitin Tests
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Protein Binding
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Saccharomyces cerevisiae Proteins / metabolism*
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Trans-Activators / metabolism
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Transcription Factors / biosynthesis*
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Transcription Factors / genetics*
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Transcription, Genetic
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Transcriptional Activation
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Transfection
Substances
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Basic Helix-Loop-Helix Transcription Factors
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DNA-Binding Proteins
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Nuclear Proteins
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Saccharomyces cerevisiae Proteins
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TCF3 protein, human
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Trans-Activators
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Transcription Factors
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Luciferases
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Acetyltransferases
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E1A-Associated p300 Protein
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Ep300 protein, mouse
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Histone Acetyltransferases