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Related Experiment Videos

Multiple positive and negative elements regulate human brain creatine kinase gene expression.

M E Ritchie1, R V Trask, H L Fontanet

  • 1Cardiovascular Division, Washington University School of Medicine, St Louis, MO 63110.

Nucleic Acids Research
|November 25, 1991
PubMed
Summary
This summary is machine-generated.

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Brain creatine kinase (BCK) gene expression rises during muscle cell development and falls in mature cells. Regulatory elements in the gene

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • The brain creatine kinase (BCK) gene plays a crucial role in cellular energy homeostasis.
  • Understanding the regulation of BCK gene expression is vital for comprehending muscle development and function.

Purpose of the Study:

  • To investigate the developmental expression pattern of the BCK gene in C2C12 myogenic cells.
  • To identify the transcriptional regulatory mechanisms governing BCK gene expression.

Main Methods:

  • Isoenzyme analysis, Western blot, and Northern blot were used to characterize BCK protein and mRNA levels.
  • Transient transfection assays with chimeric constructs of the BCK 5'-flanking DNA and the chloramphenicol acetyltransferase (CAT) reporter gene were performed.
  • Deletion analysis of the 5'-flanking region was conducted to identify regulatory elements.

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Main Results:

  • BCK subunit protein and mRNA were upregulated early in myogenesis and downregulated in differentiated myotubes.
  • Reporter gene expression from BCK 5'-flanking DNA constructs paralleled endogenous BCK mRNA levels.
  • A silencer element located between -1150 and -388 bp suppressed BCK promoter activity in both myogenic and nonmyogenic cells.

Conclusions:

  • BCK gene expression during myogenesis is controlled by sequences within its 5'-flanking DNA.
  • Negative regulatory elements in the 5'-flanking region are active in various cell types.
  • Specific elements mediate the developmental regulation of the BCK gene in muscle cells.