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Role of the CM2 Protein in Influenza C Virus Replication: Analyses of Recombinant Viruses possessing CM2 Mutants

Abstract

Yasushi Muraki *,Takako Okuwa ,Toshiki Himeda ,Yoshiro Ohara

CM2 is the second membrane protein of influenza C virus. In the present study, to investigate the role(s) of CM2 in the virus replication, we generated recombinant influenza C viruses, rC65A and rN11A, which lack the palmitoylatioin and glycosylation sites of CM2, respectively. The rC65A virus grew as efficiently as the recombinant wild-type (rWT) virus did, whereas the rN11A grew less efficiently than the rWT virus. To study the difference more precisely, we generated influenza C virus-like particles (VLPs) lacking the CM2 glycosylation site (N11A-VLPs) and examined the VLPs and VLP-infected cells. As a result, the N11A-VLPs contain approximately 13% of the virus RNA found in wildtype VLPs (WT-VLPs), and N11A-VLP-infected HMV-II cells exhibited reduced reporter gene expression compared with that of WT-VLP-infected cells (WT : N11A = 1:0.1). Thus, supportive evidence was obtained that CM2 is involved in packaging and uncoating processes and that observed growth difference between rWT and rN11A can be attributed to the difference in the CM2 function.