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Cloning and functional characterisation of avian transcription factor E2A

BACKGROUND: During B lymphocyte development the E2A gene is a critical regulator of cell proliferation and differentiation. With regards to the immunoglobulin genes the E2A proteins contribute to the regulation of gene rearrangement, expression and class switch recombination. We are now using the ch...

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Autores principales: Conlon, Thomas M, Meyer, Kerstin B
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2004
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC446191/
https://www.ncbi.nlm.nih.gov/pubmed/15196311
http://dx.doi.org/10.1186/1471-2172-5-11
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author Conlon, Thomas M
Meyer, Kerstin B
author_facet Conlon, Thomas M
Meyer, Kerstin B
author_sort Conlon, Thomas M
collection PubMed
description BACKGROUND: During B lymphocyte development the E2A gene is a critical regulator of cell proliferation and differentiation. With regards to the immunoglobulin genes the E2A proteins contribute to the regulation of gene rearrangement, expression and class switch recombination. We are now using the chicken cell line DT40 as a model system to further analyse the function of E2A. RESULTS: Here we report the cloning and functional analysis of the transcription factor E2A from chicken. Using RACE PCR on the chicken lymphoma cell line DT40 we have isolated full-length clones for the two E2A splice variants E12 and E47. Sequence conservation between the human and chicken proteins is extensive: the basic-helix-loop-helix DNA binding domain of human and chicken E47 and E12 are 93% and 92% identical, respectively. In addition high levels of conservation are seen in activation domain I, the potential NLS and the ubiquitin ligase interaction domain. E2A is expressed in a variety of tissues in chicken, with higher levels of expression in organs rich in immune cells. We demonstrate that chicken E12 and E47 proteins are strong transcriptional activators whose function depends on the presence of activation domain I. As in mammals, the dominant negative proteins Id1 and Id3 can inhibit the function of chicken E47. CONCLUSIONS: The potential for homologous recombination in DT40 allows the genetic dissection of biochemical pathways in somatic cells. With the cloning of avian E2A and the recent description of an in vitro somatic hypermutation assay in this cell line, it should now be possible to dissect the potential role of E2A in the regulation of somatic hypermutation and gene conversion.
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spelling pubmed-4461912004-07-09 Cloning and functional characterisation of avian transcription factor E2A Conlon, Thomas M Meyer, Kerstin B BMC Immunol Research Article BACKGROUND: During B lymphocyte development the E2A gene is a critical regulator of cell proliferation and differentiation. With regards to the immunoglobulin genes the E2A proteins contribute to the regulation of gene rearrangement, expression and class switch recombination. We are now using the chicken cell line DT40 as a model system to further analyse the function of E2A. RESULTS: Here we report the cloning and functional analysis of the transcription factor E2A from chicken. Using RACE PCR on the chicken lymphoma cell line DT40 we have isolated full-length clones for the two E2A splice variants E12 and E47. Sequence conservation between the human and chicken proteins is extensive: the basic-helix-loop-helix DNA binding domain of human and chicken E47 and E12 are 93% and 92% identical, respectively. In addition high levels of conservation are seen in activation domain I, the potential NLS and the ubiquitin ligase interaction domain. E2A is expressed in a variety of tissues in chicken, with higher levels of expression in organs rich in immune cells. We demonstrate that chicken E12 and E47 proteins are strong transcriptional activators whose function depends on the presence of activation domain I. As in mammals, the dominant negative proteins Id1 and Id3 can inhibit the function of chicken E47. CONCLUSIONS: The potential for homologous recombination in DT40 allows the genetic dissection of biochemical pathways in somatic cells. With the cloning of avian E2A and the recent description of an in vitro somatic hypermutation assay in this cell line, it should now be possible to dissect the potential role of E2A in the regulation of somatic hypermutation and gene conversion. BioMed Central 2004-06-14 /pmc/articles/PMC446191/ /pubmed/15196311 http://dx.doi.org/10.1186/1471-2172-5-11 Text en Copyright © 2004 Conlon and Meyer; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Research Article
Conlon, Thomas M
Meyer, Kerstin B
Cloning and functional characterisation of avian transcription factor E2A
title Cloning and functional characterisation of avian transcription factor E2A
title_full Cloning and functional characterisation of avian transcription factor E2A
title_fullStr Cloning and functional characterisation of avian transcription factor E2A
title_full_unstemmed Cloning and functional characterisation of avian transcription factor E2A
title_short Cloning and functional characterisation of avian transcription factor E2A
title_sort cloning and functional characterisation of avian transcription factor e2a
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC446191/
https://www.ncbi.nlm.nih.gov/pubmed/15196311
http://dx.doi.org/10.1186/1471-2172-5-11
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