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Packaging of Mason-Pfizer monkey virus (MPMV) genomic RNA depends upon conserved long-range interactions (LRIs)

Rawan M Kalloush1, Valérie Vivet-Boudou2, Lizna M Ali1

  • 1Department of Microbiology and Immunology, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, United Arab Emirates.

RNA (New York, N.Y.)
|April 21, 2016
PubMed

Insights

Long-range interactions (LRIs) in Mason phenylo­mye­litis virus (MPMV) genomic RNA are crucial for packaging and propagation. LRI-I is vital for structural function, while LRI-II requires more than just base-pairing.

Area of Science:

  • Molecular Biology
  • Virology
  • Gene Therapy Vectors

Background:

  • Mason phenyl­mye­litis virus (MPMV) shows promise as a gene therapy vector.
  • Understanding MPMV genomic RNA (gRNA) packaging and dimerization is critical for vector development.
  • MPMV gRNA features two conserved long-range interactions (LRIs), LRI-I and LRI-II, in its packaging signal.

Purpose of the Study:

  • To investigate the biological significance of MPMV's U5-gag LRIs in the viral life cycle.
  • To determine the roles of LRI-I and LRI-II in gRNA packaging, dimerization, and propagation.
  • To elucidate the structural requirements for LRI function in MPMV.

Main Methods:

  • Site-directed mutagenesis was used to disrupt LRI structural motifs in MPMV gRNA.
  • MPMV gRNA packaging and propagation efficiency were assessed for mutant viruses.
  • Selective 2'hydroxyl acylation analyzed by primer extension (SHAPE) was employed to probe RNA structure.
  • In vitro RNA dimerization assays were performed to evaluate dimerization capabilities.

Main Results:

  • Disruption of LRI base-pairing significantly impaired MPMV gRNA packaging and propagation.
  • A double mutant with a restored heterologous LRI-I was fully functional, indicating its structural role.
  • A similar LRI-II mutant failed to restore function, suggesting base-pairing alone is insufficient for LRI-II.
  • Impaired RNA packaging in LRI mutants was not due to defects in RNA dimerization.

Conclusions:

  • MPMV U5-gag LRIs are essential for gRNA packaging and viral propagation.
  • LRI-I primarily functions at a structural level, maintaining gRNA architecture.
  • LRI-II function is more complex than simple base-pairing, requiring specific structural contributions.
  • These findings highlight the architectural importance of LRIs in the 5' region of MPMV gRNA, extending their known roles to nonlentiviral retroviruses.
Keywords:
Mason-Pfizer monkey virus (MPMV)RNA packaging and dimerizationRNA secondary structureSHAPE (selective 2′hydroxyl acylation analyzed by primer extension)long-range interactions (LRI)retroviruses

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