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Related Experiment Videos

Identification of eukaryotic mRNAs that are translated at reduced cap binding complex eIF4F concentrations using a

G Johannes1, M S Carter, M B Eisen

  • 1Department of Microbiology, Stanford University School of Medicine, Stanford, CA 94305, USA.

Proceedings of the National Academy of Sciences of the United States of America
|November 11, 1999
PubMed
Summary

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Many cellular mRNAs can bypass the cap binding complex eIF4F for translation initiation. This internal ribosome entry mechanism allows translation of key stress response genes even when eIF4F is depleted.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Virology

Background:

  • Most eukaryotic messenger RNAs (mRNAs) require the cap binding complex eIF4F for cap-dependent translation initiation.
  • Alternative internal ribosome entry mechanisms allow translation of certain viral and cellular RNAs when eIF4F levels are low.

Purpose of the Study:

  • To identify cellular mRNAs that utilize internal ribosome entry sites (IRES) for translation when eIF4F is depleted.
  • To investigate the role of IRES in stress response gene expression.

Main Methods:

  • Human cells infected with poliovirus were used to deplete eIF4F.
  • Messenger RNAs associated with polysomes under low eIF4F conditions were isolated and identified using cDNA microarray.
  • Internal ribosome entry sites in selected mRNAs were confirmed through functional assays.

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Main Results:

  • Approximately 200 out of 7000 analyzed mRNAs remained associated with polysomes when eIF4F was depleted.
  • These mRNAs encoded proteins involved in transcription, signaling pathways (MAPK), protooncogenes (c-myc, Pim-1), and angiogenesis (Cyr61).
  • Internal ribosome entry sites were confirmed in Cyr61 and Pim-1 mRNAs.

Conclusions:

  • Diverse cellular mRNAs, particularly those involved in stress responses like inflammation and angiogenesis, can employ cap-independent translation initiation.
  • This IRES-mediated translation is crucial for maintaining the expression of essential genes under conditions of eIF4F scarcity.