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Methods to Discover Alternative Promoter Usage and Transcriptional Regulation of Murine Bcrp1
Published on: May 27, 2016
Structure of the Mesothelin/MPF gene and characterization of its promoter
1University Department of Medicine, University of Western Australia, Queen Elizabeth II Medical Centre, 4th Floor, G Block, Nedlands, Perth, Western Australia, 6009, Australia.
Abstract:
Mesothelin is a cell-surface antigen of unknown function that is expressed in a highly tissue-specific manner. Expression of the protein is limited to mesothelium, the tissue forming the pleural, pericardial, and peritoneal membranes. The purpose of this study was to characterize the gene and identify elements responsible for its transcription. The gene contains an 1884-bp open reading frame encoded by 15 exons occupying 8 kb of human chromosome 16. An 1850-bp region of genomic DNA at the 5' end of the gene encompassing the proposed transcriptional start site was cloned. This region lacks a TATA box and other regulatory elements such as SP1 sites, which are commonly found in promoters. Transient transfection analyses demonstrated that mesothelium-specific control elements are present within the 1.85-kb region. Minimal constitutive promoter elements were localized to a 317-bp region. Tissue-specific enhancer elements upstream of the minimal promoter were found to activate transcription from the homologous and a heterologous promoter in a position- and orientation-independent manner.
Insights
Researchers characterized the mesothelin gene, identifying regulatory elements responsible for its tissue-specific transcription. This discovery sheds light on the control mechanisms governing mesothelin gene expression in mesothelial cells.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- Mesothelin is a cell-surface antigen with an unknown function.
- Its expression is highly tissue-specific, primarily in mesothelium (pleural, pericardial, peritoneal membranes).
Purpose of the Study:
- To characterize the mesothelin gene.
- To identify DNA elements regulating mesothelin gene transcription.
Main Methods:
- Cloning of the 5' genomic region of the mesothelin gene.
- Transient transfection analyses to assess promoter and enhancer activity.
Main Results:
- The mesothelin gene contains a 1884-bp open reading frame across 15 exons.
- The 5' flanking region lacks typical promoter elements like TATA boxes and SP1 sites.
- A 1.85-kb region contains mesothelium-specific control elements.
- Minimal constitutive promoter elements are within a 317-bp region.
- Upstream enhancer elements activate transcription independently of position and orientation.
Conclusions:
- The identified regulatory elements are crucial for mesothelium-specific mesothelin gene expression.
- Understanding these elements provides insights into the transcriptional control of mesothelin.
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