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https://hdl.handle.net/2440/95555
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Type: | Journal article |
Title: | A NF-kappa B/Sp1 region is essential for chromatin remodeling and correct transcription of a human granulocyte-macrophage colony-stimulating factor transgene |
Author: | Cakouros, D. Cockerill, P.N. Bert, A.G. Mital, R. Roberts, D.C. Shannon, M.F. |
Citation: | Journal of Immunology, 2001; 167(1):302-310 |
Publisher: | American Association of Immunologists |
Issue Date: | 2001 |
ISSN: | 1550-6606 0022-1767 |
Statement of Responsibility: | Dimitrios Cakouros, Peter N. Cockerill, Andrew G. Bert, Renu Mital, Donna C. Roberts, and M. Frances Shannon |
Abstract: | The GM-CSF gene is expressed following activation of T cells. The proximal promoter and an upstream enhancer have previously been characterized using transfection and reporter assays in T cell lines in culture. A 10.5-kb transgene containing the entire human GM-CSF gene has also been shown to display inducible, position-independent, copy number-dependent transcription in mouse splenocytes. To determine the role of individual promoter elements in transgene function, mutations were introduced into the proximal promoter and activity assessed following the generation of transgenic mice. Of four mutations introduced into the transgene promoter, only one, in an NF-kappaB/Sp1 region, led to decreased induction of the transgene in splenocytes or bone marrow-derived macrophages. This mutation also affected the activity of reporter gene constructs stably transfected into T cell lines in culture, but not when transiently transfected into the same cell lines. The mutation alters the NF-kappaB family members that bind to the NF-kappaB site as well as reducing the binding of Sp1 to an adjacent element. A DNase I hypersensitive site that is normally generated at the promoter following T cell activation on the wild-type transgene does not appear in the mutant transgene. These results suggest that the NF-kappaB/Sp1 region plays a critical role in chromatin remodeling and transcription on the GM-CSF promoter in primary T cells. |
Keywords: | Cells, Cultured Jurkat Cells Chromatin Animals Mice, Inbred C57BL Mice, Inbred CBA Mice, Transgenic Humans Mice NF-kappa B Granulocyte-Macrophage Colony-Stimulating Factor DNA Transfection Mutagenesis, Site-Directed Transcription, Genetic Gene Expression Regulation Response Elements Transgenes Sp1 Transcription Factor Promoter Regions, Genetic CD28 Antigens |
Rights: | Copyright © 2001 by The American Association of Immunologists |
DOI: | 10.4049/jimmunol.167.1.302 |
Published version: | http://dx.doi.org/10.4049/jimmunol.167.1.302 |
Appears in Collections: | Aurora harvest 7 Medicine publications |
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