Identification of Saint Louis encephalitis virus mRNA

C. W. Naeve, D. W. Trent

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Saint Louis encephalitis (SLE) virus-specific RNA was recovered from infected HeLa cells by sodium dodecyl sulfate (SDS)-phenol-chloroform extraction, and the molecular species were resolved by SDS-sucrose gradient centrifugation and agarose gel electrophoresis. Sucrose gradient centrifugation revealed the presence of a 45S species, minor 20 to 30S heterogeneous species, and an 8 to 10S RNA species in the cytoplasmic extract. Analysis of the same samples by electrophoresis on agarose gels, under both nondenaturing and denaturing conditions, revealed only two virus-specific RNA molecules, the 45S genome-sized RNA and an 8 to 10S species. Varying the gel concentration to facilitate analysis of nucleic acids with molecular weights ranging from 25,000 to 25 x 106 failed to reveal additional RNA species, although low levels of a putative double-stranded replicative form could conceivably have escaped detection. From our observations it appears that the heterogeneous RNA species and presumably the 20S RNase-resistant species reported in other investigations of flavivirus RNA are degradation products or conformers of the 45S molecule. Polysomes from SLE virus-infected cells were prepared and separated from contaminating nucleocapsid by centrifugation on discontinuous sucrose gradients. RNA extracted from these polysome preparations was analyzed by sucrose gradient centrifugation and agarose gel electrophoresis. The 45S SLE virus genome-size molecule was found to be the only RNA species associated with the polysomes. This molecule was sensitive to RNase digestion and was released from polysomes by EDTA and puromycin treatment. These findings provide direct evidence that the 45S SLE virus RNA serves as the messenger during virus replication, in contrast to the 26S RNA species which functions as the predominant messenger during alphavirus replication.

Original languageEnglish (US)
Pages (from-to)535-545
Number of pages11
JournalJournal of Virology
Volume25
Issue number2
StatePublished - 1978

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St. Louis Encephalitis Viruses
Saint Louis encephalitis virus
RNA
Messenger RNA
Polyribosomes
Centrifugation
polyribosomes
Sucrose
Agar Gel Electrophoresis
centrifugation
sucrose
agarose
Ribonucleases
Sodium Dodecyl Sulfate
sodium dodecyl sulfate
gel electrophoresis
Alphavirus
Flavivirus
Genome Size
Nucleocapsid

ASJC Scopus subject areas

  • Immunology

Cite this

Naeve, C. W., & Trent, D. W. (1978). Identification of Saint Louis encephalitis virus mRNA. Journal of Virology, 25(2), 535-545.

Identification of Saint Louis encephalitis virus mRNA. / Naeve, C. W.; Trent, D. W.

In: Journal of Virology, Vol. 25, No. 2, 1978, p. 535-545.

Research output: Contribution to journalArticle

Naeve, CW & Trent, DW 1978, 'Identification of Saint Louis encephalitis virus mRNA', Journal of Virology, vol. 25, no. 2, pp. 535-545.
Naeve, C. W. ; Trent, D. W. / Identification of Saint Louis encephalitis virus mRNA. In: Journal of Virology. 1978 ; Vol. 25, No. 2. pp. 535-545.
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