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Human Tumor Necrosis Factor-α Gene 3ⲠUntranslated Region Confers Inducible Toxin Responsiveness to Homologous Promoter in Monocytic THP-1 Cells
To better define the role of 3â² untranslated region (3â²UTR) on transcriptional regulation of the human tumor necrosis factor (TNF)-α gene, monocytic human THP-1 cells were transfected with two TNF-α promoter constructs spanning base pairs â1897/â1 and â1214/â1, respectively, and linked...
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Published in: | The Journal of biological chemistry 1999-07, Vol.274 (31), p.21714 |
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Main Authors: | , , |
Format: | Article |
Language: | English |
Online Access: | Get full text |
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Summary: | To better define the role of 3â² untranslated region (3â²UTR) on transcriptional regulation of the human tumor necrosis factor
(TNF)-α gene, monocytic human THP-1 cells were transfected with two TNF-α promoter constructs spanning base pairs â1897/â1
and â1214/â1, respectively, and linked to the rabbit β-globin gene. Quantitative globin gene expression of chimerae was measured
by reverse transcription-polymerase chain reaction. A construct linking the chicken β-actin promoter and a deleted portion
of the β-globin gene was cotransfected and used as internal standard. Unexpectedly, when THP-1 cells were stimulated with
lipopolysaccharide or toxic shock syndrome toxin-1, gene regulation was hardly detected. In contrast, endogenous TNF-α gene
regulation measured by the same reverse transcription-polymerase chain reaction procedure was vigorous. Remarkably, ligation
of 3â²UTR to chimeric constructs led to a drastic drop in the basal level of chimeric gene expression, resulting in a 15- to
40-fold induction of the reporter gene. Consistently, when the TNF-α promoter was replaced by the cytomegalovirus early immediate
promoter, gene expression was also uniformly reduced but was no longer up-regulated upon stimulation with lipopolysaccharide
and toxic shock syndrome toxin-1. These data provide the first line of evidence that, in addition to its role in TNF-α transcript
stability and translation, human TNF-α 3â²UTR also participates in modulating gene expression at the transcriptional level. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.274.31.21714 |