The avian reovirus genome segment S1 is a functionally tricistronic gene that expresses one structural and two nonstructural proteins in infected cells

The avian reovirus S1 gene contains three partially overlapping, out-of-phase open reading frames (ORFs) that are highly conserved in all avian reovirus strains examined to date. The three S1 ORFs of the avian reovirus strain S1133 were individually expressed in bacterial cells, and their purified translation products used as antigens to raise specific polyclonal antibodies. With these antibodies we were able to demonstrate that all three S1 ORFs from different avian reovirus strains are translatable in infected cells. Proteins p10 and p17, which are specified by ORF1 and ORF2, respectively, are nonstructural proteins which associate with cell membranes, whereas ORF3 directs the synthesis of protein sigmaC, a structural oligomeric protein responsible for cell attachment. While intracellular synthesis of protein sigmaC was demonstrated a long time ago and that of protein p10 was reported recently, this is the first time that expression of the S1 ORF2 has been demonstrated experimentally, Thus, the previously reported coding capacity of the avian reovirus genome is now expanded to 14 proteins, of which ten are structural (lambdaA, lambdaB, lambdaC, muA, muB, mu BC, mu BN, sigmaA, sigmaB, and sigmaC) and four are nonstructural (mu NS, sigma NS, p17, and p10). Finally, protein p10, but not p17 or sigmaC, induces cell-cell fusion when transiently expressed in mammalian cells, supporting a previously published observation that the polypeptide encoded by the S1 ORF1 plays an important role in the syncytial phenotype displayed by avian reoviruses. (C) 2001 Academic Press.