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Presence of prepackaged mRNA in virions of DNA adenovirus

Siu-Wah Chung1, John A Arnott, Yizeng Yang

  • 1Department of Pathology & Laboratory Medicine, Fels Institute for Cancer Research & Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA.

The Journal of Biological Chemistry
|October 3, 2003
PubMed

Insights

Ran GTPase influences innate immunity. Adenovirus (Ad) vector administration showed distinct tissue distribution in mice, particularly in macrophages, due to pre-packaged viral mRNA enabling immediate protein translation.

Area of Science:

  • Immunology
  • Molecular Biology
  • Virology

Background:

  • Ran GTPase plays a role in host innate immune responses.
  • Two Ran alleles, RanT/n and RanC/d, exhibit opposing effects on immunity.
  • Adenovirus (Ad) vectors are utilized for gene transfer, but their tissue distribution can vary.

Purpose of the Study:

  • To investigate the impact of Ran alleles on host innate immune response.
  • To analyze the tissue distribution of Ad vectors following different administration routes.
  • To explore the role of pre-packaged viral mRNA in early gene expression and vector distribution.

Main Methods:

  • Adenovirus (Ad) vectors carrying different Ran alleles were administered intravenously and intraperitoneally in mice.
  • Vector tissue distribution was assessed using real-time PCR.
  • Presence and function of pre-packaged RNA within Ad virions and infected cells were analyzed using actinomycin-D and real-time PCR.

Main Results:

  • Intraperitoneal administration of Ad vectors resulted in effective transduction into peritoneal macrophages and distinct tissue distribution patterns within 2 hours.
  • Pre-packaged viral mRNA was detected within Ad virions and infected cells, with approximately one copy per viral genome.
  • Experiments using actinomycin-D confirmed that this pre-packaged mRNA could be translated into proteins.

Conclusions:

  • Immediate translation of Ran proteins from pre-packaged viral mRNA after Ad vector uncoating in the cytoplasm is a key mechanism for early differences in vector tissue distribution.
  • This mechanism contributes to efficient gene transfer into macrophages and influences the host innate immune response.

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