Smooth muscle calponin: An unconventional CArG-dependent gene that antagonizes neointimal formation

Xiaochun Long, Orazio J. Slivano, Sarah L. Cowan, Mary A. Georger, Ting Hein Lee, Joseph M. Miano

Research output: Contribution to journalArticle

Abstract

OBJECTIVE-: Smooth muscle calponin (CNN1) contains multiple conserved intronic CArG elements that bind serum response factor and display enhancer activity in vitro. The objectives here were to evaluate these CArG elements for activity in transgenic mice and determine the effect of human CNN1 on injury-induced vascular remodeling. METHODS AND RESULTS-: Mice carrying a lacZ reporter under control of intronic CArG elements in the human CNN1 gene failed to show smooth muscle cell (SMC)-restricted activity. However, deletion of the orthologous sequences in mice abolished endogenous Cnn1 promoter activity, suggesting their necessity for in vivo Cnn1 expression. Mice carrying a 38-kb bacterial artificial chromosome (BAC) harboring the human CNN1 gene displayed SMC-restricted expression of the corresponding CNN1 protein, as measured by immunohistochemistry and Western blotting. Extensive BAC recombineering studies revealed the absolute necessity of a single intronic CArG element for correct SMC-restricted expression of human CNN1. Overexpressing human CNN1 suppressed neointimal formation following arterial injury. Mice with an identical BAC carrying mutations in CArG elements that inhibit human CNN1 expression showed outward remodeling and neointimal formation. CONCLUSION-: A single intronic CArG element is necessary but insufficient for proper CNN1 expression in vivo. CNN1 overexpression antagonizes arterial injury-induced neointimal formation.

Original languageEnglish (US)
Pages (from-to)2172-2180
Number of pages9
JournalArteriosclerosis, thrombosis, and vascular biology
Volume31
Issue number10
DOIs
StatePublished - Oct 1 2011
Externally publishedYes

Fingerprint

Smooth Muscle
Bacterial Artificial Chromosomes
Genes
Smooth Muscle Myocytes
Wounds and Injuries
Serum Response Factor
Sequence Deletion
Transgenic Mice
calponin
Western Blotting
Immunohistochemistry
Mutation
Proteins

Keywords

  • gene expression
  • genetically altered mice
  • serum response factor
  • transgenic models
  • vascular muscle

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Smooth muscle calponin : An unconventional CArG-dependent gene that antagonizes neointimal formation. / Long, Xiaochun; Slivano, Orazio J.; Cowan, Sarah L.; Georger, Mary A.; Lee, Ting Hein; Miano, Joseph M.

In: Arteriosclerosis, thrombosis, and vascular biology, Vol. 31, No. 10, 01.10.2011, p. 2172-2180.

Research output: Contribution to journalArticle

Long, Xiaochun ; Slivano, Orazio J. ; Cowan, Sarah L. ; Georger, Mary A. ; Lee, Ting Hein ; Miano, Joseph M. / Smooth muscle calponin : An unconventional CArG-dependent gene that antagonizes neointimal formation. In: Arteriosclerosis, thrombosis, and vascular biology. 2011 ; Vol. 31, No. 10. pp. 2172-2180.
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