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Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle-specific expression of a single contractile protein gene.

Publication ,  Journal Article
Lee, KJ; Ross, RS; Rockman, HA; Harris, AN; O'Brien, TX; van Bilsen, M; Shubeita, HE; Kandolf, R; Brem, G; Price, J
Published in: J Biol Chem
August 5, 1992

To examine the relationship between the cardiac and skeletal muscle gene programs, the current study employs the regulatory (phosphorylatable) myosin light chain (MLC-2) as a model system. Northern blotting, primer extension, and RNase protection studies documented the high level expression of the cardiac MLC-2 mRNA in both mouse cardiac and slow skeletal muscle (soleus). Transgenic mouse lines harboring a 2100- or a 250-base pair rat cardiac MLC-2 promoter/luciferase fusion gene were generated, demonstrating high levels of luciferase activity in cardiac muscle, and only background luminescence in slow skeletal muscle and non-muscle tissues. As assessed by in situ hybridization, immunofluorescence, and luminescence assays of luciferase reporter activity in various regions of the heart, both the endogenous MLC-2 gene and the MLC-2 luciferase fusion gene were expressed exclusively in the ventricular compartment, with expression in the atrium at background levels. Point mutations within the conserved regulatory sites HF-1a and HF-1b significantly cripple ventricular muscle specificity, while mutation of the single E-box site was without effect, suggesting that ventricular muscle-specific expression occurs through an E-box-independent pathway. This study provides direct evidence that the cis regulatory sequences in the cardiac/slow twitch MLC-2 gene which confer cardiac and skeletal muscle-specific expression can be clearly segregated, suggesting that distinct regulatory programs may have evolved to control the tissue-specific expression of this single contractile protein gene in cardiac and skeletal muscle.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

August 5, 1992

Volume

267

Issue

22

Start / End Page

15875 / 15885

Location

United States

Related Subject Headings

  • Sequence Homology, Nucleic Acid
  • Recombinant Fusion Proteins
  • RNA
  • Organ Specificity
  • Oligodeoxyribonucleotides
  • Myosins
  • Muscles
  • Molecular Sequence Data
  • Mice, Transgenic
  • Mice
 

Citation

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Lee, K. J., Ross, R. S., Rockman, H. A., Harris, A. N., O’Brien, T. X., van Bilsen, M., … Price, J. (1992). Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle-specific expression of a single contractile protein gene. J Biol Chem, 267(22), 15875–15885.
Lee, K. J., R. S. Ross, H. A. Rockman, A. N. Harris, T. X. O’Brien, M. van Bilsen, H. E. Shubeita, R. Kandolf, G. Brem, and J. Price. “Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle-specific expression of a single contractile protein gene.J Biol Chem 267, no. 22 (August 5, 1992): 15875–85.
Lee KJ, Ross RS, Rockman HA, Harris AN, O’Brien TX, van Bilsen M, et al. Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle-specific expression of a single contractile protein gene. J Biol Chem. 1992 Aug 5;267(22):15875–85.
Lee KJ, Ross RS, Rockman HA, Harris AN, O’Brien TX, van Bilsen M, Shubeita HE, Kandolf R, Brem G, Price J. Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle-specific expression of a single contractile protein gene. J Biol Chem. 1992 Aug 5;267(22):15875–15885.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

August 5, 1992

Volume

267

Issue

22

Start / End Page

15875 / 15885

Location

United States

Related Subject Headings

  • Sequence Homology, Nucleic Acid
  • Recombinant Fusion Proteins
  • RNA
  • Organ Specificity
  • Oligodeoxyribonucleotides
  • Myosins
  • Muscles
  • Molecular Sequence Data
  • Mice, Transgenic
  • Mice