The c-rel protooncogene product c-Rel but not NF-κB binds to the intronic region of the human interferon-γ gene at a site related to an interferon-stimulable response element

Antonio Sica, Tse Hua Tan, Nancy Rice, Marcus Kretzschmar, Paritosh Ghosh, Howard A. Young

Research output: Contribution to journalArticle

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Abstract

Interferon-γ (IFN-γ) is an important immunoregulatory protein that is expressed usually only in large granular lymphocytes and T cells. The gene encoding IFN-γ was previously found to contain an intronic enhancer element that was not tissue-specific in its activity, despite the restricted expression of the intact IFN-γ-encoding gene. Using nuclear extracts from the human T-cell line Jurkat, we have now identified two protein-binding regions in this intronic enhancer element. One of the protected regions has strong partial identity to the NF-κB site present in the promoter region of the human interleukin 2-encoding gene. Based on this observation and recent reports of the interaction of the c-rel protooncogene product (c-Rel) with NF-κB sites, we determined whether c-Rel could interact with the intronic enhancer element in the human IFN-γ genomic DNA. Most surprisingly, gel-shift analysis, using c-Rel expressed in Escherichia coli established that c-Rel binds specifically to the IFN-γ intronic DNA but not to the interleukin 2-like NF-κB site. Additional studies with antibodies prepared against c-Rel peptides verified specificity of the interaction of c-Rel with this binding site. In addition, using an affinity-purified p50 subunit of the NF-κB complex, we observed that the p50 protein did not bind to this additional c-Rel-binding site. Furthermore, nucleotide sequence analysis of this DNA region revealed a strong similarity of the additional c-Rel-binding site to a previously identified IFN-stimulable response element. These data show that c-Rel can interact with DNA regions distinct from that recognized by NF-κB and may, in fact, be involved in transcriptional regulation of the IFN-stimulable genes via the IFN-stimulable response element.

Original languageEnglish
Pages (from-to)1740-1744
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number5
Publication statusPublished - 1992

Fingerprint

Response Elements
Interferons
Genes
DNA
Binding Sites
Interleukin-2
T-Lymphocytes
Electrophoretic Mobility Shift Assay
Genetic Promoter Regions
Protein Binding
Sequence Analysis
Proteins
Lymphocytes
Escherichia coli
Cell Line
Peptides
Antibodies

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

The c-rel protooncogene product c-Rel but not NF-κB binds to the intronic region of the human interferon-γ gene at a site related to an interferon-stimulable response element. / Sica, Antonio; Tan, Tse Hua; Rice, Nancy; Kretzschmar, Marcus; Ghosh, Paritosh; Young, Howard A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 5, 1992, p. 1740-1744.

Research output: Contribution to journalArticle

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abstract = "Interferon-γ (IFN-γ) is an important immunoregulatory protein that is expressed usually only in large granular lymphocytes and T cells. The gene encoding IFN-γ was previously found to contain an intronic enhancer element that was not tissue-specific in its activity, despite the restricted expression of the intact IFN-γ-encoding gene. Using nuclear extracts from the human T-cell line Jurkat, we have now identified two protein-binding regions in this intronic enhancer element. One of the protected regions has strong partial identity to the NF-κB site present in the promoter region of the human interleukin 2-encoding gene. Based on this observation and recent reports of the interaction of the c-rel protooncogene product (c-Rel) with NF-κB sites, we determined whether c-Rel could interact with the intronic enhancer element in the human IFN-γ genomic DNA. Most surprisingly, gel-shift analysis, using c-Rel expressed in Escherichia coli established that c-Rel binds specifically to the IFN-γ intronic DNA but not to the interleukin 2-like NF-κB site. Additional studies with antibodies prepared against c-Rel peptides verified specificity of the interaction of c-Rel with this binding site. In addition, using an affinity-purified p50 subunit of the NF-κB complex, we observed that the p50 protein did not bind to this additional c-Rel-binding site. Furthermore, nucleotide sequence analysis of this DNA region revealed a strong similarity of the additional c-Rel-binding site to a previously identified IFN-stimulable response element. These data show that c-Rel can interact with DNA regions distinct from that recognized by NF-κB and may, in fact, be involved in transcriptional regulation of the IFN-stimulable genes via the IFN-stimulable response element.",
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