Adenovirus-mediated transfer of the smooth muscle cell calponin gene inhibits proliferation of smooth muscle cells and fibroblasts

Zhong Jiang, Robert W. Grange, Michael P. Walsh, Kristine E. Kamm

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Smooth muscle cell calponin (h1 or basic isoform) is an actin-binding protein that inhibits actomyosin MgATPase activity and is abundantly expressed in differentiated smooth muscle. Western blots showed bovine tracheal (BT) smooth muscle cells in culture expressed only 2±1% (n=8) of the amount of calponin in tissues, while NIH-3T3 fibroblasts expressed none. We tested the hypothesis that introduction of calponin to cultured BT and 3T3 cells would inhibit cytoskeletal activities associated with cell proliferation. To achieve high-efficiency expression, an adenovirus encoding the CMV-calponin construct (Adv-CaP) was generated by homologous recombination in 293 cells. With greater than 90% of BT and 3T3 cells infected with Adv-CaP, calponin expression (32 and 11 μg/mg total protein, respectively) was similar to that in smooth muscle tissues (51 μg/mg). Cells were infected with Adv-CaP for 48 h, replated at low density and proliferation rates were assessed by cell density and [3Hlthymidine incorporation. Cell growth and DNA synthesis by Adv-CaP-infected cells were inhibited to one-third control values for both BT and 3T3 cells. Expressed calponin was localized primarily on stress fibers in both cell types. Calponin may act at the cytoskeletal level to retard signaling pathways that normally lead to tight coupling between cell shape and DNA synthesis.

Original languageEnglish (US)
Pages (from-to)441-445
Number of pages5
JournalFEBS Letters
Volume413
Issue number3
DOIs
StatePublished - Aug 25 1997

Fingerprint

Fibroblasts
Adenoviridae
Smooth Muscle Myocytes
Muscle
Genes
Cells
3T3 Cells
Smooth Muscle
Tissue
Microfilament Proteins
Actomyosin
Stress Fibers
Cell Shape
Homologous Recombination
DNA
Cell proliferation
Cell growth
calponin
Cell culture
Protein Isoforms

Keywords

  • Actin-binding protein
  • Adenovirus
  • Calponin
  • Fibroblast
  • Smooth muscle

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Adenovirus-mediated transfer of the smooth muscle cell calponin gene inhibits proliferation of smooth muscle cells and fibroblasts. / Jiang, Zhong; Grange, Robert W.; Walsh, Michael P.; Kamm, Kristine E.

In: FEBS Letters, Vol. 413, No. 3, 25.08.1997, p. 441-445.

Research output: Contribution to journalArticle

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