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Cis-regulatory elements of the mouse Krt1.12 gene
I-Jong Wang1, Eric C Carlson, Chia-Yang Liu
1Department of Ophthalmology, University of Cincinnati, Cincinnati, OH, USA. Winston.Kao@uc.edu
Molecular Vision
|April 16, 2002
Summary
Researchers identified specific DNA regions upstream of the Krt1.12 gene that regulate cornea-specific keratin 12 expression. These cis-regulatory elements involve interactions of transcription factors like AP1, c/EBP, and KLF6.
Area of Science:
- Molecular biology
- Genetics
- Ophthalmology
Background:
- Keratin 12 is crucial for cornea epithelial cell structure and function.
- Understanding the regulation of Keratin 12 expression is key to cornea biology.
Purpose of the Study:
- To identify cis-regulatory elements controlling Keratin 12 gene expression.
- To investigate the DNA sequences between the transcription start site and 600 bp upstream of the Krt1.12 gene.
Main Methods:
- Reporter gene constructs (0.6, 0.4, 0.2 kb) of the Krt1.12 promoter were tested in rabbit corneas using Gene Gun technology.
- DNA footprinting and electrophoretic mobility shift assays (EMSA) were performed with bovine corneal epithelial cell nuclear extracts.
- Bioinformatic analysis identified potential transcription factor binding sites.
Main Results:
- Elements between -0.2 and -0.6 kb upstream of Krt1.12 drive corneal epithelial cell-specific expression.
- Specific DNA regions (-181 to -111 and -256 to -193) bound nuclear proteins.
- These regions contain putative binding sites for transcription factors including AP1, c/EBP, and KLF6.
Conclusions:
- The DNA fragments -182 to -111 and -256 to -193 function as cornea-specific cis-regulatory elements.
- Coordinated action of multiple transcription factors is necessary for cornea-specific Krt1.12 gene expression.