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Related Experiment Videos

Murine muscle-specific enolase: cDNA cloning, sequence, and developmental expression.

N Lamandé1, A M Mazo, M Lucas

  • 1Laboratoire de Biochimie Cellulaire, Collège de France, Paris.

Proceedings of the National Academy of Sciences of the United States of America
|June 1, 1989
PubMed
Summary

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Researchers cloned the beta subunit of mouse muscle-specific enolase, finding its expression is regulated by mRNA levels during muscle development. This suggests beta enolase may mark the determination of myogenic lineage.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Developmental Biology

Background:

  • Vertebrates express glycolytic enzyme enolase as homo- and heterodimers of alpha, beta, and gamma subunits.
  • Skeletal muscle development involves a transition from alpha alpha to beta beta enolase forms.

Purpose of the Study:

  • To clone and sequence the cDNA for the beta subunit of murine muscle-specific enolase.
  • To investigate the regulation of alpha and beta enolase mRNA levels during muscle development.
  • To determine if beta enolase expression is an early marker of myogenic lineage determination.

Main Methods:

  • Cloning and sequencing of beta-enolase cDNA from mouse.
  • Amino acid sequence homology comparison with chicken beta-enolase and rat/mouse alpha and gamma enolases.

Related Experiment Videos

  • Analysis of 3' untranslated regions of mouse alpha, beta, and gamma enolase mRNAs.
  • Study of enolase gene expression in myogenic cell lines.
  • Transfection experiments using MyoD1 cDNA in premyogenic cells.

    • Main Results:

    • The cloned beta-enolase amino acid sequence shows >80% homology with chicken beta-enolase and other mouse/rat enolase subunits.
    • Mouse alpha, beta, and gamma enolase mRNAs exhibit significant differences in their 3' untranslated regions and lengths.
    • The transition to beta beta enolase in developing muscle is primarily due to differential regulation of alpha and beta mRNA levels.
    • Beta enolase gene expression begins at the myoblast stage.
    • Initial beta transcript expression occurs early in the myogenic pathway upon MyoD1 introduction.

    Conclusions:

    • The beta subunit of murine muscle-specific enolase has been cloned and sequenced.
    • Differential regulation of alpha and beta enolase mRNA levels controls the subunit composition during muscle development.
    • Early beta enolase expression suggests its potential role as a marker for myogenic lineage determination.