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A regulatory element within a coding exon modulates keratin 18 gene expression in transgenic mice
N Neznanov1, A Umezawa, R G Oshima
1Burnham Institute, La Jolla, California 92037, USA.
The Journal of Biological Chemistry
|November 5, 1997
Summary
The keratin 18 (K18) gene
Area of Science:
- Molecular Biology
- Gene Regulation
- Epigenetics
Background:
- Tissue-specific DNase-hypersensitive sites are linked to regulatory regions of the human keratin 18 (K18) gene.
- Exon 6 of the K18 gene contains a potential AP-1 binding site, interacting with c-Jun and c-Fos in vitro.
Purpose of the Study:
- To investigate the regulatory role of K18 exon 6 in gene expression.
- To determine the impact of mutations within K18 exon 6 on gene regulation and expression levels.
Main Methods:
- Reporter gene assays in F9 cells.
- Analysis of transgenic mice.
- Transient transfections of mutant K18 gene constructs.
- Footprint analysis for protein binding.
Main Results:
- K18 exon 6 enhances reporter gene expression and modulates AP-1 responsiveness.
- Mutations in exon 6 decreased but did not abolish regulatory activity, maintaining tissue specificity.
- A frameshift mutation in exon 6 drastically reduced K18 expression (<10% of wild type) due to decreased RNA stability and a novel negative regulatory element.
Conclusions:
- The protein-coding region of the K18 gene, specifically exon 6, possesses intrinsic regulatory functions.
- Exon 6 plays a significant role in K18 gene expression, RNA stability, and interaction with other regulatory elements.