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Irreversible steps in the ferritin synthesis induction pathway

L S Goessling1, D P Mascotti, M Bhattacharyya-Pakrasi

  • 1Department of Biology, Washington University, St. Louis, Missouri 63130.

The Journal of Biological Chemistry
|February 11, 1994
PubMed
Summary

Cellular ferritin synthesis re-repression is slow and requires new protein synthesis. Iron or heme induction of ferritin synthesis appears irreversible due to slow, irreversible covalent modification of IRE-binding protein (IRE-BP).

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Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Ferritin synthesis is regulated by iron and heme levels.
  • The regulation involves the iron-responsive element-binding protein (IRE-BP).
  • The precise mechanisms of ferritin synthesis re-repression and its reversibility are not fully understood.

Purpose of the Study:

  • To investigate the kinetics and requirements for re-repression of ferritin synthesis after iron or heme removal.
  • To determine if covalent modification of IRE-BP contributes to the irreversibility of ferritin induction.

Main Methods:

  • Cell culture experiments measuring ferritin synthesis rates.
  • Treatment with iron, heme, Desferrioxamine mesylate (Desferal), and cycloheximide.
  • Analysis of IRE-binding protein (IRE-BP) modification and regeneration using Western blotting or similar techniques.

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Main Results:

  • Re-repression of ferritin synthesis is a slow process, taking 4-10 hours.
  • New protein synthesis is required for re-repression, as indicated by cycloheximide inhibition.
  • Ferritin synthesis induction is largely irreversible without an iron chelator.
  • Heme- or iron-dependent IRE-BP modification occurs rapidly but is slowly reversed.
  • Prolonged exposure to heme leads to irreversible IRE-BP modification, correlating with ferritin induction irreversibility.

Conclusions:

  • Ferritin synthesis re-repression is a slow, protein synthesis-dependent process.
  • The induction of ferritin synthesis is essentially irreversible in the absence of iron chelation.
  • Slow, irreversible covalent modification of IRE-BP is a key mechanism underlying the persistent induction of ferritin synthesis, particularly in rapidly growing cells.