RNA and polypeptide homology among murine coronaviruses

Abstract

Using a =P complementary DNA (cDNA) prepared against the A59 nucleocapsid protein messenger RNA, we have investigated the extent of homology between A59 and four other strains of murine hepatitis virus (MHV). Analysis by hybridization kinetics of the annealing between A59 [S2P]cDNA and infected cell RNA from the other four MHV strains demonstrated 7040% homology. By gel transfer analysis, the A59 [S2P]eDNA was able to detect subgenomic-size virus-specific RNAs in cells infected with all of the five MHV strains. A similar pattern of seven viral RNAs was detected in cells infected with A59, MHV-1, MHV-3, and JHM. In contrast, cells infected with MHV-S contained seven virusspecific RNAs, of which only the two smallest species comigrated with RNAs from the other four strains. The results suggest that as previously shown with A59 (S. Cheley, R. Anderson, M. J. Cupples, E. C. M. Lee Chan, and V. L. Morris (1981) Virdogy, 112,596-604), all MHV strains tested encode a nested set of subgenomic RNAs. Analysis of [?l]methionine-labeled viral proteins by SDS-polyacrylamidegel electrophoresis revealed that each strain of MHV specified four major viral polypeptides with apparent molecular weights very similar to those previously reported for the E2, N, El, and PEl polypeptides of A59. The strong degree of interstrain homology among the five MHV strains investigated was confirmed by comparative chymotryptic peptide mapping of the viral N proteins. A majority of the chymotryptic peptides from each of the [35S$nethioninelabeled N proteins was found to coelute by high-performance liquid chromatography. Moreover, this technique of peptide mapping indicated a particularly strong relatedness between MHV-1 and MHV-S and among MHV3, JHM, and A59.

Antigenic relationships of murine coronaviruses: Analysis using monoclonal antibodies to JHM (MHV-4) virus

“…Although these strains have not all been previously compared in a single study, previous investigations do support this general division . For example, the oligonucleotide maps of MHV-1 and MHV-S are distinct from the other MHVs examined , as are the peptide maps of the pp6O from these two strains (Cheley et at, 1981) . In addition, the oligonucleotide maps of A59, MHV-3, MHV-D, and MHV-K are very similar Lai, unpublished data) .…”

“…Nevertheless, the members of the first group characteristically produce other diseases in addition to hepatitis: A59 can cause demyelination choroidoependymitis (Virelizier et at, 1975) MHV-D enteritis (Ishida et at, 1978a), and MHV-K myeloproliferation in nude mice (Ishida et at,197Sb) . By contrast, the members of the second group (MHV-2, MHV-S, and MHV-1) have been reported to produce primarily hepatitis ; MHV-1 and MHV-S have also been considered to be of relatively low pathogenicity (Cheley et at, 1981;.…”

“…Most MHV strains are antigenically distinct by these methods, although the MHV3 and A59 strains are very similar. Analysis of the viral polypeptides has shown that some MHVs have proteins with unique migration patterns by sodium dodecyl sulfate polyacrylamide gel electrophoresis Cheley et at, 1981; . More recently, several studies have compared the oligonueleotide maps of closely related MHVs that cause different diseases in order to identify the genes responsible for different disease patterns Wege et al, 1981 ;Stohlman et at ., 1982) .…”

SARS: Lessons Learned from Other Coronaviruses

“…Coronaviruses are 60-200 nm round, somewhat pleiomorphic, particles containing an internal helical RNA-protein nucleocapsid surrounded by an envelope containing glycoprotein peplomers (15) (Fig. 1).…”

Viruses, Immunity, and Mental Disorders

“…Such Northern blots reveal significant differences between the RNAs present during pennissive and latent infections. Under pennissive conditions, all seven mRNAs (Cheley et al, 1981) of JHMV are detected (Fig. 2).…”