Sequence and expression of the ns2 protein gene of human coronavirus OC43

Abstract

The complete nucleotide sequence of the ns2 gene of human coronavirus OC43 (HCV-OC43) was determined. Sequence analysis revealed an open reading frame that could encode a protein of 278 amino acids, with an estimated molecular mass of 32.2 kDa. Six potential phosphorylation sites are present but no sites of N-glycosylation were found. The amino acid sequence of the HCV-OC43 ns2 protein shows 92 % identity with that of the Mebus strain of bovine coronavirus (BCV). However, a stretch of nine consecutive amino acids near the C terminus is completely different, causing it to be very hydrophilic, which contrasts with the hydrophobic nature of this region in BCV. As shown by immunofluorescence with a monospecific antiserum, the ns2 protein was expressed in the cytoplasm of HCV-OC43-infected HRT-18 cells.

The Group-Specific Murine Coronavirus Genes Are Not Essential, but Their Deletion, by Reverse Genetics, Is Attenuating in the Natural Host

“…No enzymatic activities have been assigned to the other group-specific proteins. The 31-kDa 2a protein was detected in the cytoplasm of MHV, bovine coronavirus (BCoV), and human coronavirus (HCoV)-OC43 infected cells (Bredenbeek et al, 1990a;Cox et al, 1991;Labonte et al, 1995;Zoltick et al, 1990). The MHV and BCoV 2a proteins share 45% amino acid identity.…”

Human Papillomavirus Type 16 E7 Peptide-Directed CD8+T Cells from Patients with Cervical Cancer Are Cross-Reactive with the Coronavirus NS2 Protein

“…Additional target cell lines included the HLA-A2 ϩ , HPV16 ϩ cervical cancer cell line Caski (38,42) or the HLA-A2 ϩ B-cell line C1R-A2 (see below). C1R-A2 cells were either transfected with the entire coronavirus OC43 NS2 gene (25) to test for recog-nition of naturally processed and presented peptides or with the melanomaassociated target antigen NY-ESO1 (22) as a control in the CMV-driven TA expression vector (Invitrogen, Groningen, The Netherlands). Peptides (listed in Table 1) were pulsed onto T2 cells plus human ␤ 2 -microglobulin to test for peptide-specific T-cell recognition.…”

Human Respiratory Coronavirus OC43: Genetic Stability and Neuroinvasion

“…The PCR products encompassed the entire genome of the viruses and overlapped each other to make sure the final sequences were complete. Primers used for the amplifications of ORF 1a and ORF1b were created by using the sequence of the bovine coronavirus Quebec strain (BCoV Quebec) (63), which displays 97% identity in this region of the genome and was known to share 92% identity with the 3Ј 9 kb of the HCoV-OC43 genome (24,26,29,37,38,39). Primers used to amplify the 3Ј region were designed on the basis of sequences of HCoV-OC43 available in GenBank.…”