Polyadenylate in the Virion RNA of Mouse Hepatitis Virus

Abstract

Mouse hepatitis (MH) virus was grown in SR-CDF1-DBT, a mouse cell line, and purified by ammonium sulfate precipitation and by density gradient centrifugation. Extraction of RNA from purified virions with 1% SDS and sedimentation analysis of the RNA revealed a major 50S component and two minor components. Treatment of virions with phenol/chloroform also produced the 50S component, although its yield was lower. MH virion RNA can bind to a poly(U)-fiberglass filter, indicating that MH virion RNA contains poly(A). A poly(A)-like fragment was isolated by digestion with ribonuclease A [EC 3.1.4.22] and T1 [EC 3.1.4.8] and by DEAE-Sephadex column chromatography. Analysis of the fragment for base composition showed it to be an adenine-rich material. Its chain length was about 90 nucleotides, as determined by ion-exchange chromatography and gel electrophoresis.

The virus-specific intracellular RNA species of two murine coronaviruses: MHV-A59 and MHV-JHM

“…Poly(A) tracts are seen as streaks in the upper left hand corner of the fingerprints of A59V and JHMV virion RNAs. This confirms the findings of others (Yogo et al, 1977;Lai and Stohlman, 1978;Wege et al, 1978) that the MHV genome is polyadenylated. The seven major A59V-and JHMV-specific intracellular RNAs are also polyadenylated.…”

“…Coronaviruses are defined as pleomorphic enveloped particles about 100 nm in diameter which have characteristic bul- bous projections on their surface, bud through the endoplasmic reticulum rather than the plasma membrane, and contain RNA (Tyrell et al, 1978). Several studies of coronavirus genomic RNA have been reported (Lomneczi, 197'7;Lomneczi and Kennedy, 1977;Yogo et al, 1977;Lai and Stohlman, 1978;Macnaughton and Madge, 1978;Schochetman et al, 1977;Tannock and Hierholzer, 1978;Guy and Brian, 1979;Wege et al, 1978). These studies have indicated that the coronavirus genome is a large (5.4-8.1 X lo6 daltons) singlestranded RNA which is polyadenylated and is infectious.…”

Pathogenic murine coronaviruses I. Characterization of biological behavior in vitro and virus-specific intracellular RNA of strongly neurotropic JHMV and weakly neurotropic A59V viruses

“…Lai and Stohlman (1978) have shown that the virions of A59V and JHMV contain a single-stranded RNA with a molecular weight of 5.4 x lo6 daltons. The virion RNAs of MHV-2 (Yogo et al, 1977), JHMV, and A59V (Lai and Stohlman, 1978) contain tracts of polyadenylic acid. The viron RNA of avian infectious bronchitis virus, an avian coronavirus, contains at least one tract of polyadenylic acid and is infectious (Schochetman et al, 1977, Lomniczi, 1977.…”

Analysis of genomic and intracellular viral RNAs of small plaque mutants of mouse hepatitis virus, JHM strain

“…Present address: Department of Microbiology, School of Medicine, University of Southern California, 2925 Zonal Avenue, HMR-401, Los Angeles, Calif. 99933. *Present address: Institute of Virology and Immunobiology, University of Wiirzburg, Versbacher Strasse '7,8'709 Wiirzburg,Federal Republic of Germany. 1981; Wege et al, 1978Wege et al, , 1981Yogo et aL, 1977). In cells infected with MHV six species of subgenomic mRNA, together with a genomic-size mRNA, are produced (and are numbered in order of decreasing size).…”