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The 5'CL-PCBP RNP complex, 3' poly(A) tail and 2A(pro) are required for optimal translation of poliovirus RNA

Sushma A Ogram1, Allyn Spear, Nidhi Sharma

  • 1Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610-0245, USA.

Virology
|December 1, 2009
PubMed

Insights

Optimal translation of poliovirus RNA requires the 5' cap-binding protein (5'CL-PCBP) complex, 3' poly(A) tail, and viral protein 2A(pro). These elements independently and synergistically enhance viral RNA translation.

Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • Poliovirus RNA translation is a complex process regulated by viral and cellular factors.
  • The 5' cap-binding protein (5'CL-PCBP) complex and the 3' poly(A) tail are known to play roles in viral RNA translation.
  • The viral protein 2A(pro) is a protease implicated in viral replication and translation regulation.

Purpose of the Study:

  • To elucidate the specific roles of the 5'CL-PCBP complex, 3' poly(A) tail, and viral protein 2A(pro) in poliovirus RNA translation.
  • To determine the functional domain of PCBP involved in viral RNA translation.
  • To investigate the interplay between these elements in regulating translation efficiency.

Main Methods:

  • Protein-RNA tethering assays were employed to study the interactions between viral RNA elements and proteins.
  • Translation efficiency was measured in the presence or absence of specific viral components.
  • Site-directed mutagenesis was used to identify the functional domain of PCBP.

Main Results:

  • The 5'CL-PCBP complex, 3' poly(A) tail, and viral protein 2A(pro) are all essential for optimal poliovirus RNA translation.
  • The KH3 domain of PCBP2 was identified as the critical functional region within the 5'CL-PCBP complex.
  • The 5'CL-PCBP complex and 3' poly(A) tail independently stimulate translation, but also function synergistically to enhance and prolong translation.
  • 2A(pro)-mediated translation stimulation occurs irrespective of the presence of the 5'CL and 3' poly(A) tail.

Conclusions:

  • A model is proposed where the 5'CL-PCBP complex interacts with the 3' poly(A)-PABP complex to form a 5'-3' circular structure.
  • This circularization facilitates ribosome reloading, thereby enhancing poliovirus RNA translation.
  • The findings highlight a sophisticated mechanism for regulating viral translation through coordinated interactions of RNA elements and viral proteins.

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