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Related Experiment Videos

Cyclic AMP-responsive element binding protein in brain mitochondria.

M Cammarota1, G Paratcha, L R Bevilaqua

  • 1Instituto de Biologia Celular y Neurociencias Prof. Dr. Eduardo de Robertis, Facultad de Medicina, Universidad de Buenos Aires, Argentina.

Journal of Neurochemistry
|June 1, 1999
PubMed
Summary

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Cyclic AMP-responsive element binding protein (CREB) is found in rat brain mitochondria, not just the nucleus. This discovery suggests CREB may play a role in mitochondrial function and cellular signaling.

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Cell Biology

Background:

  • Cyclic AMP-responsive element binding protein (CREB) is crucial for brain functions like memory formation.
  • CREB is a well-known transcription factor and a nuclear target of signal transduction pathways.
  • Its role outside the nucleus, particularly in mitochondria, is largely unexplored.

Purpose of the Study:

  • To investigate the subcellular localization of CREB in the rat brain.
  • To determine if CREB is present and functional within mitochondria.
  • To explore potential roles of mitochondrial CREB in cellular signaling and gene expression.

Main Methods:

  • Subcellular fractionation of rat hippocampus and cerebral cortex.
  • Immunoreactivity assays for phosphorylated CREB (pCREB) in mitochondrial fractions.

Related Experiment Videos

  • In vitro phosphorylation and dephosphorylation assays of mitochondrial CREB.
  • DNA mobility shift assays using mitochondrial extracts.
  • Immunoelectron microscopy to pinpoint pCREB localization.

    • Main Results:

    • CREB, including its phosphorylated form (pCREB), was unexpectedly detected in both synaptic and nonsynaptic mitochondria from rat brain.
    • Mitochondrial CREB can be phosphorylated by protein kinase A and dephosphorylated by protein phosphatases.
    • Mitochondrial extracts exhibit CREB-binding activity to a specific DNA element, confirmed by antibody supershift assays.
    • Immunoelectron microscopy localized pCREB to the inner mitochondrial membrane.

    Conclusions:

    • CREB is not exclusively a nuclear protein but is also localized within brain mitochondria.
    • Mitochondrial CREB is biochemically active and can be regulated by phosphorylation/dephosphorylation.
    • These findings suggest a novel role for CREB in mitochondrial function and the integration of extracellular signals with gene expression.