Nanoparticle encapsidation of Flock House virus by auto assembly of Tobacco Mosaic virus coat protein

Payal D. Maharaj, Jyothi K. Mallajosyula, Gloria Lee, Phillip Thi, Yiyang Zhou, Christopher M. Kearney, Alison A. McCormick

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Tobacco Mosaic virus (TMV) coat protein is well known for its ability to self-assemble into supramolecular nanoparticles, either as protein discs or as rods originating from the ~300 bp genomic RNA origin-of-assembly (OA). We have utilized TMV self-assembly characteristics to create a novel Flock House virus (FHV) RNA nanoparticle. FHV encodes a viral polymerase supporting autonomous replication of the FHV genome, which makes it an attractive candidate for viral transgene expression studies and targeted RNA delivery into host cells. However, FHV viral genome size is strictly limited by native FHV capsid. To determine if this packaging restriction could be eliminated, FHV was adapted to express enhanced green fluorescent protein (GFP), to allow for monitoring of functional FHV RNA activity. Then TMV OA was introduced in six 3' insertion sites, with only site one supporting functional FHV GFP expression. To create nanoparticles, FHV GFP-OA modified genomic RNA was mixed in vitro with TMV coat protein and monitored for encapsidation by agarose electrophoresis and electron microscopy. The production of TMV-like rod shaped nanoparticles indicated that modified FHV RNA can be encapsidated by purified TMV coat protein by self-assembly. This is the first demonstration of replication-independent packaging of the FHV genome by protein self-assembly.

Original languageEnglish (US)
Pages (from-to)18540-18556
Number of pages17
JournalInternational Journal of Molecular Sciences
Volume15
Issue number10
DOIs
StatePublished - Oct 14 2014
Externally publishedYes

Fingerprint

Tobacco Mosaic Virus
Virus Assembly
tobacco
Tobacco
Capsid Proteins
viruses
Viruses
Nanoparticles
assembly
proteins
Proteins
nanoparticles
RNA Viruses
RNA
Product Packaging
Green Fluorescent Proteins
genome
Genome
Self assembly
Genome Size

Keywords

  • Encapsidation
  • Flock House virus
  • Nanoparticle
  • Self-assembly
  • Tobacco Mosaic virus
  • Transgene expression
  • Viral vector

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Spectroscopy
  • Inorganic Chemistry
  • Catalysis
  • Molecular Biology
  • Computer Science Applications
  • Medicine(all)

Cite this

Nanoparticle encapsidation of Flock House virus by auto assembly of Tobacco Mosaic virus coat protein. / Maharaj, Payal D.; Mallajosyula, Jyothi K.; Lee, Gloria; Thi, Phillip; Zhou, Yiyang; Kearney, Christopher M.; McCormick, Alison A.

In: International Journal of Molecular Sciences, Vol. 15, No. 10, 14.10.2014, p. 18540-18556.

Research output: Contribution to journalArticle

Maharaj, Payal D. ; Mallajosyula, Jyothi K. ; Lee, Gloria ; Thi, Phillip ; Zhou, Yiyang ; Kearney, Christopher M. ; McCormick, Alison A. / Nanoparticle encapsidation of Flock House virus by auto assembly of Tobacco Mosaic virus coat protein. In: International Journal of Molecular Sciences. 2014 ; Vol. 15, No. 10. pp. 18540-18556.
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