Source:http://linkedlifedata.com/resource/pubmed/id/17023546
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
2007-2-9
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| pubmed:databankReference | |
| pubmed:abstractText |
Growing evidence that epithelial CD98 plays an important role in intestinal inflammation focused our interest to investigate the transcriptional regulation of CD98. Our mouse-based in vivo and in vitro experiments revealed that epithelial colonic CD98 mRNA expression was transcriptionally increased in intestinal inflammation. We then isolated and characterized a 5'-flanking fragment containing the promoter region required for CD98 gene transcription. Primer extension and rapid amplification of 5'-cDNA ends were used to map a transcriptional initiation site 129 bp upstream from the translational start codon (ATG). Direct sequencing of the 5'-flanking region revealed the presence of four GC-rich stimulating protein (Sp)1 binding domains, one NF-kappaB binding domain, and no TATA box. Binding of Sp1 [Sp1(-874), SP1(-386), Sp1(-187), and Sp1(-177)] and NF-kappaB [NF-kappaB(-213)] to the promoter was confirmed by EMSA and supershift assays. Furthermore, chromatin immunoprecipitation experiments showed the in vivo DNA-Sp1 and DNA-NF-kappaB interactions under basal and IFN-gamma-stimulated conditions. Reporter genes driven by serially truncated and site-mutated CD98 promoters were used to examine basal and IFN-gamma-responsive transcription in transiently transfected Caco2-BBE cells. Our results revealed that Sp1(-187), Sp1(-177), and the NF-kappaB binding site were essential for basal and IFN-gamma-stimulated CD98 promoter activities, whereas Sp1(-874) and Sp1(-386) were not. The results from additional site-mutated CD98 promoters suggested that Sp1(-187), Sp1(-177), and the NF-kappaB site may cooperate in mediating basal and IFN-gamma-stimulated CD98 promoter activities. Finally, we demonstrated that a reduction of Sp1 or NF-kappaB expression reduced CD98 protein expression in unstimulated and IFN-gamma-stimulated Caco2-BBE cells. Collectively, these findings indicate that the Sp1 and NF-kappaB transcription factors are likely to play a significant role in IFN-gamma-mediated transcriptional regulation of CD98 in the intestinal epithelium, providing new insights into the regulation of CD98 expression in intestinal inflammation.
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| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, CD98,
http://linkedlifedata.com/resource/pubmed/chemical/Interferon-gamma,
http://linkedlifedata.com/resource/pubmed/chemical/Luciferases,
http://linkedlifedata.com/resource/pubmed/chemical/NF-kappa B,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Small Interfering,
http://linkedlifedata.com/resource/pubmed/chemical/Sp1 Transcription Factor
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| pubmed:status |
MEDLINE
|
| pubmed:month |
Feb
|
| pubmed:issn |
0193-1857
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
292
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| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
G535-45
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| pubmed:dateRevised |
2008-11-21
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| pubmed:meshHeading |
pubmed-meshheading:17023546-Animals,
pubmed-meshheading:17023546-Antigens, CD98,
pubmed-meshheading:17023546-Base Sequence,
pubmed-meshheading:17023546-Binding Sites,
pubmed-meshheading:17023546-Caco-2 Cells,
pubmed-meshheading:17023546-Chromatin Immunoprecipitation,
pubmed-meshheading:17023546-Colitis,
pubmed-meshheading:17023546-Electrophoretic Mobility Shift Assay,
pubmed-meshheading:17023546-Female,
pubmed-meshheading:17023546-Gene Expression Regulation,
pubmed-meshheading:17023546-Humans,
pubmed-meshheading:17023546-Interferon-gamma,
pubmed-meshheading:17023546-Intestinal Mucosa,
pubmed-meshheading:17023546-Luciferases,
pubmed-meshheading:17023546-Mice,
pubmed-meshheading:17023546-Mice, Inbred C57BL,
pubmed-meshheading:17023546-Molecular Sequence Data,
pubmed-meshheading:17023546-Mutation,
pubmed-meshheading:17023546-NF-kappa B,
pubmed-meshheading:17023546-Promoter Regions, Genetic,
pubmed-meshheading:17023546-Protein Binding,
pubmed-meshheading:17023546-RNA, Small Interfering,
pubmed-meshheading:17023546-Sp1 Transcription Factor,
pubmed-meshheading:17023546-Transcription Initiation Site,
pubmed-meshheading:17023546-Transfection
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| pubmed:year |
2007
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| pubmed:articleTitle |
Characterization of the human intestinal CD98 promoter and its regulation by interferon-gamma.
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| pubmed:affiliation |
Div of Digestive Diseases, Dept of Medicine, Emory Univ, Atlanta, GA 30322, USA.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
|