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A retinoic acid-responsive element in human midkine gene
C Pedraza1, S Matsubara, T Muramatsu
1Department of Biochemistry, Faculty of Medicine, Kagoshima University.
Journal of Biochemistry
|April 1, 1995
Summary
Researchers identified a specific DNA sequence in the human Midkine (MK) gene that responds to retinoic acid. This element is crucial for regulating gene expression, with slight variations noted compared to mouse and consensus sequences.
Area of Science:
- Molecular Biology
- Gene Regulation
- Developmental Biology
Background:
- Midkine (MK) is a growth/differentiation factor.
- MK is a product of a retinoic acid-responsive gene.
Purpose of the Study:
- To identify cis-acting elements in the human MK gene's upstream sequence responsible for retinoic acid-induced expression.
- To characterize the retinoic acid-responsive element (RARE) within the human MK gene.
Main Methods:
- Cloning the 2.3-kb upstream sequence of the human MK gene.
- Constructing fusion genes with chloramphenicol acetyl-transferase (CAT) reporter gene.
- Transfecting F9 embryonal carcinoma cells and measuring CAT activity after retinoic acid treatment.
- Performing deletion analysis of the identified cis-acting element.
Main Results:
- The 2.3-kb upstream sequence conferred retinoic acid-induced expression of the CAT gene.
- A DR5-type RARE, characterized by direct repeats, was identified in the 5'-region of the human MK gene.
- Deletion of this direct repeat significantly reduced retinoic acid-induced CAT gene expression.
- The core RARE sequence in the human gene showed minor nucleotide differences from the consensus RARE and the mouse MK gene RARE.
Conclusions:
- A specific retinoic acid-responsive element in the human MK gene regulates its expression.
- This element plays a critical role in mediating retinoic acid's effect on MK gene transcription.
- Sequence variations in the RARE may influence the precise regulation of Midkine expression across species.