The Genome of Human Coronavirus Strain 229E

Abstract

SUMMARYThe genomic RNA of human coronavirus strain 229E (HCV 229E) migrated on polyacrylamide gels as a single peak with a tool. wt. of 5"8 x IO 6. Denaturation of the genome with formaldehyde did not alter its electrophoretic mobility, which suggests that the I-ICV Ee9E genome is a single-stranded molecule. At least 3o ~ of the genomic RNA was shown to contain covalently attached polyadenylic acid [poly(A)] sequences by binding the RNA to an oligo(dT)-cellulose column. These poly(A) tracts were shown to be about 7o nucleotides in length by measuring the resistance to digestion of HCV 229E RNA with pancreatic and T1 RNases. Finally, the genomic RNA was shown to terminate at or near the 3'-terminus on the basis of its susceptibility to polynucleotide phosphorylase.

The Biology of Coronaviruses

“…of 5 × 106 to 7 × 10 6, corresponding to about 15000 to 20000 nucleotides. Ta-resistant oligonucleotide fingerprinting of genome RNA and intracellular viral mRNA (see Coronavirus-directed RNA synthesis) confirms the positive polarity of the genome and indicates that it does not have extensive sequence reiteration (Brian et al, 1980;Clewley et al, 1981 ;King & Brian, 1982;Lai & Stohlman, 1978Lai et al, 1981 ;Leibowitz et al, 1981;Macnaughton, 1978;Macnaughton & Madge, 1978;Spaan et al, 1981Spaan et al, , 1982Stern & Kennedy, 1980a, b;Wege et al, 1978Wege et al, , 1981aWeiss & Leibowitz, 1981).…”

Serological Relationships of the Subcomponents of Human Coronavirus Strain 229E and Mouse Hepatitis Virus Strain 3

“…The reactions were significantly above the level with preimmune sera, although the heterologous reactions were lower than the homologous reactions. Antisera to HCV 229E and MHV 3 RNPs did not react by ELISA against purified HCV 229E RNA isolated as described previously (Macnaughton & Madge, 1978), indicating that the cross-reactions were not due to RNA components of RNP. ELISAs of antisera to envelope against HCV 229E (Fig.…”

“…HCV 229E and MHV 3 were grown and purified as previously described (Macnaughton & Madge, 1978;. Virus structural components comprising surface projections, envelope and RNP were prepared from purified, disrupted virus particles and separated on sucrose density gradients as described previously Macnaughton et al, 1981), Subcomponents were shown to be uncontaminated with the other components by labelling their polypeptides with 1251 and analysing them on polyacrylamide gels Macnaughton et al, 1981).…”

Enzyme-linked Immunosorbent Assay for Coronaviruses HCV 229E and MHV 3

“…The virus strains used and the preparation of purified virus suspensions for biochemical and electron microscopic analysis have been described previously (Macnaughton & Madge, 1978;Macnaughton et al I978). HCV 229E was grown in monolayer cultures of human cells of the MRC continuous (MRCc) line at 33 °C for 32 h in Eagle's BME with 2 °/o newborn calf serum (Flow Laboratories, Irvine, Scotland) or with Lt5 medium with 2 % rabbit serum -these were called antigens HCV 229E (N) and HCV 229E (R) respectively.…”