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Related Experiment Videos

A cell cycle-dependent protein serves as a template-specific translation initiation factor.

E V Pilipenko1, T V Pestova, V G Kolupaeva

  • 1Institute of Poliomyelitis and Viral Encephalitides, Russian Academy of Medical Sciences, Moscow 142782, Russia.

Genes & Development
|August 19, 2000
PubMed
Summary

Cell-specific translation control is mediated by IRES-binding proteins like ITAF(45). These RNA chaperones, such as ITAF(45) and PTB, dictate picornavirus IRES function and cell-type specificity.

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Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Cap-independent translation initiation in picornaviruses relies on internal ribosomal entry sites (IRES) within the 5' UTR.
  • This process requires eukaryotic initiation factors (eIFs) and IRES-trans-acting factors (ITAFs).

Purpose of the Study:

  • To investigate how differences in ITAFs contribute to cell type-specific IRES function among picornaviruses.
  • To elucidate the molecular mechanisms underlying IRES-mediated translation control.

Main Methods:

  • Substitution of the Theiler's murine encephalomyelitis virus (TMEV) IRES with the foot-and-mouth disease virus (FMDV) IRES.
  • Reconstitution of translation initiation using fractionated cellular components.
  • Chemical and enzymatic footprinting to map protein-binding sites on the FMDV IRES.

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Main Results:

  • The FMDV IRES, unlike the TMEV IRES, requires the proliferation-dependent protein ITAF(45) (murine proliferation-associated protein, Mpp1) for efficient 48S complex formation.
  • ITAF(45) is not expressed in murine brain cells, explaining the attenuation of TMEV neurovirulence when its IRES is replaced by the FMDV IRES.
  • PTB and ITAF(45) function cooperatively as RNA chaperones, promoting the stable binding of eIF4G/4A to the FMDV IRES.

Conclusions:

  • Differences in ITAF requirements explain cell type-specific IRES activity.
  • The distribution of ITAFs, such as PTB and ITAF(45), contributes to the cell-specific translational control of viral and potentially cellular mRNAs.
  • ITAFs act as critical regulators modulating the functional state of specific IRES elements.