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Negative control elements and cAMP responsive sequences in the tissue-specific expression of mouse renin genes
1Molecular and Cellular Vascular Research Laboratory, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
Summary
Mouse renin genes Ren1d and Ren2d have regulatory elements controlling their expression. A 160-base-pair insertion in Ren2d disrupts a negative control element, affecting gene regulation.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- The 5' flanking regions of mouse renin genes (Ren1d and Ren2d) possess potential regulatory elements.
- Sequence analysis revealed a 160-base-pair insertion in the Ren2d gene's regulatory region.
Purpose of the Study:
- To investigate the functional roles of the 5' flanking regions of mouse Ren1d and Ren2d genes.
- To determine the impact of the insertion in Ren2d on gene regulation.
Main Methods:
- Cloning of 5' flanking regions fused to a chloramphenicol acetyltransferase (CAT) reporter gene under a thymidine kinase (TK) promoter.
- Transfection of chimeric constructs into AtT-20 and JEG-3 cells.
- Assessing CAT expression under basal and cAMP-stimulated conditions.
Main Results:
- The Ren1d 5' flanking sequence inhibited basal CAT expression in the sense orientation.
- The Ren2d 5' flanking region showed no basal inhibition, indicating a non-functional negative control element.
- Both Ren1d and Ren2d demonstrated a 3-fold increase in transcription in response to 8-bromo-cAMP, confirming a functional cAMP-responsive element.
Conclusions:
- The 160-base-pair insertion in Ren2d likely interferes with the negative control element's function.
- The cAMP-responsive element is functional in both mouse renin genes.
- These findings contribute to understanding the tissue-specific regulation of mouse renin genes.