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Intronic CArG Box Regulates Cysteine-Rich Protein 2 Expression in the Adult but Not in Developing Vasculature

Intronic CArG Box Regulates Cysteine-Rich Protein 2 Expression in the Adult but Not in Developing... Intronic CArG Box Regulates Cysteine-Rich Protein 2 Expression in the Adult but Not in Developing Vasculature Chung-Huang Chen, Meng-Ling Wu, Yi-Chung Lee, Matthew D. Layne, Shaw-Fang Yet Objective—An absence of cysteine-rich protein 2 (CRP2) enhances vascular smooth muscle cell (VSMC) migration and increases neointima formation after arterial injury; therefore, CRP2 plays an important role in the response to vascular injury. The goal of the present study was to elucidate the molecular mechanisms that preserve CRP2 expression in the adult vasculature and thus might serve to inhibit the response to injury. Methods and Results—We generated a series of transgenic mice harboring potential Csrp2 regulatory regions with a lacZ reporter. We determined that the 12-kb first intron was necessary for transgene activity in adult but not in developing vasculature. Within the intron we identified a 6.3-kb region that contains 2 CArG boxes. Serum response factor preferentially bound to CArG2 box in gel mobility shift and chromatin immunoprecipitation assays; additionally, serum response factor coactivator myocardin factors activated CRP2 expression via the CArG2 box. Mutational analysis revealed that CArG2 box was important in directing lacZ expression in VSMC of adult vessels. Conclusion—Although CRP2 expression during development is independent of CArG box regulatory sites, http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Arteriosclerosis, Thrombosis, and Vascular Biology Wolters Kluwer Health

Intronic CArG Box Regulates Cysteine-Rich Protein 2 Expression in the Adult but Not in Developing Vasculature

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/lp/wolters-kluwer-health/intronic-carg-box-regulates-cysteine-rich-protein-2-expression-in-the-MqsmIrZS4C
ISSN
1079-5642
eISSN
1524-4636
DOI
10.1161/ATVBAHA.109.197251
pmid
20075421

Abstract

Intronic CArG Box Regulates Cysteine-Rich Protein 2 Expression in the Adult but Not in Developing Vasculature Chung-Huang Chen, Meng-Ling Wu, Yi-Chung Lee, Matthew D. Layne, Shaw-Fang Yet Objective—An absence of cysteine-rich protein 2 (CRP2) enhances vascular smooth muscle cell (VSMC) migration and increases neointima formation after arterial injury; therefore, CRP2 plays an important role in the response to vascular injury. The goal of the present study was to elucidate the molecular mechanisms that preserve CRP2 expression in the adult vasculature and thus might serve to inhibit the response to injury. Methods and Results—We generated a series of transgenic mice harboring potential Csrp2 regulatory regions with a lacZ reporter. We determined that the 12-kb first intron was necessary for transgene activity in adult but not in developing vasculature. Within the intron we identified a 6.3-kb region that contains 2 CArG boxes. Serum response factor preferentially bound to CArG2 box in gel mobility shift and chromatin immunoprecipitation assays; additionally, serum response factor coactivator myocardin factors activated CRP2 expression via the CArG2 box. Mutational analysis revealed that CArG2 box was important in directing lacZ expression in VSMC of adult vessels. Conclusion—Although CRP2 expression during development is independent of CArG box regulatory sites,

Journal

Arteriosclerosis, Thrombosis, and Vascular BiologyWolters Kluwer Health

Published: Apr 1, 2010

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