Mouse hepatitis virus A59: mRNA structure and genetic localization of the sequence divergence from hepatotropic strain MHV-3

Abstract

The composition and structure of the mouse hepatitis virus (MHV)-specific RNA in actinomycin D-treated, infected L-2 cells were studied. SEven virus-specific RNA species with molecular weights of 0.6 X 10(6), 0.9 X 10(6), 1.2 X 10(6), 1.5 X 10(6), 3.0 X 10(6), 4.0 X 10(6), and 5.4 X 10(6) (equivalent to the viral genome) were detected. T1 oligonucleotide fingerprinting studies suggested that the sequences of each RNA species were totally included within the next large RNa species. The oligonucleotides of each RNA species were mapped on the 60S RNA genome of the virus. Each RNA species contained the oligonucleotides starting from the 3' end of the genome and extending continuously for various lengths in the 3' leads to 5' direction. All of the viral RNA species contained a polyadenylate stretch of 100 to 130 nucleotides and probably identical sequences immediately next to the polyadenylate. These data suggested that the virus-specific RNAs are mRNA's and have a stairlike structure similar to that of infectious bronchitis virus, an avian coronavirus. A proposal is presented, based on the mRNA structure, for the designation of the genes on the MHV genome. Using this proposal, the sequence differences between A59, a weakly pathogenic strain, and MHV-3, a strongly hepatotropic strain, were localized primarily in mRNA's 1 and 3, corresponding t genes A and C.

Characterization of N protein self-association in coronavirus ribonucleoprotein complexes

“…The plaque-cloned A59 strain of MHV (MHV-A59) (Lai et al, 1981) and its temperature-sensitive mutant Alb4 (Koetzner et al, 1992) were used for this study. MHV-A59 and Alb4 stock viruses were propagated in mouse DBT cells (Hirano et al, 1974) at 37 • C and 33 • C, respectively.…”

“…The M and E proteins are essential for viral envelope formation and release (Bos et al, 1996;Vennema et al, 1996). In infected cells, the virus produces an intracellular form of genomic RNA, mRNA 1, and six to eight species of subgenomic mRNAs (Lai et al, 1981;Leibowitz et al, 1981;Stern and Kennedy, 1980a,b). These virus-specific mRNAs comprise a nested set with a common 3 -terminus (Lai et al, 1981;Leibowitz et al, 1981;Stern and Kennedy, 1980a,b) and a common leader sequence of approximately 60-100 nucleotides at the 5 -end (Lai and Cavanagh, 1997).…”

Two novel neutralizing antigenic epitopes of the s1 subunit protein of a QX-like avian infectious bronchitis virus strain Sczy3 as revealed using a phage display peptide library

“…IBV is an enveloped, non-segmented, single-stranded, positivesense RNA virus with a genome of approximately 27.6 kb (Boursnell et al, 1987). The remaining one-third of the genome encodes four main structural proteins: the spike glycoprotein (S), the small membrane protein (E), the integral membrane protein (M), and the nucleocapsid protein (N) (Lai et al, 1981). The S glycoprotein is a large type I transmembrane glycoprotein that is responsible for receptor binding and membrane fusion (Cavanagh et al, 1986).…”

Importance of coronavirus negative-strand genomic RNA synthesis prior to subgenomic RNA transcription

“…The MHV-specific genes, which are all located downstream from the leader, are separated from one another by an intergenic region. MHV-infected cells produce genomic-size mRNA and six to seven species of subgenomic mRNA species that make up a 3%-coterminal nested group, in which each mRNA carries one more gene in the 5% direction than the last (Lai et al, 1981;Leibowitz et al, 1981). All these virusspecific mRNAs have a leader RNA sequence at their 5%-end (Lai et al, , 1984Spaan et al, 1983) and a poly (A) tail at the 3%-end.…”