Independent loss events of a functional tetherin gene in galliform birds

ABSTRACT The restriction factor tetherin (bone marrow stromal cell antigen 2) is an interferon-inducible protein preventing the release of newly formed viral particles from infected cells. Tetherin displays antiviral activity against a broad range of enveloped viruses, including retroviruses. While tetherin orthologs have been identified in several mammalian species, little is known about its expression and activity in non-mammalian vertebrates, including birds. We have previously described antiviral activity of chicken ( Gallus gallus ) tetherin against the prototypical avian retrovirus avian sarcoma and leukosis virus (ASLV). Here, we report the loss of functional tetherin orthologs in several galliform birds, including turkey ( Meleagris gallopavo ) and Mikado pheasant ( Syrmaticus mikado ). In both species, the tetherin coding sequence acquired inactivating mutations, including an in-frame stop codon and frameshifting deletions. Similar to the chicken tetherin ortholog, reconstituted turkey and Mikado pheasant tetherins restricted ASLV and HIV-1 indicating antiviral activity of the ancestor protein. Previous work revealed the presence of the TMCC(aT ) gene in close proximity to tetherin, encoding a protein with tetherin-like structure in multiple vertebrates. Intriguingly, ectopic overexpression of chicken and turkey TMCC(aT) proteins in human cells decreased total HIV-1 yield. In contrast to tetherin, however, TMCC(aT) did not specifically restrict virion release, suggesting distinct antiviral mechanisms. In line with this, IFNα stimulation did not reduce the release of ASLV particles from infected turkey cells. Overall, our data describe the loss of functional tetherin genes in several galliform species and identify an antiviral activity of the related TMCC(aT) protein that is mechanistically different from that of tetherin. IMPORTANCE Birds represent important hosts for numerous viruses, including zoonotic viruses and pathogens with the potential to cause major economic losses to the poultry industry. Viral replication and transmission can be inhibited or blocked by the action of antiviral restriction factors (RFs) encoded by the host. One well-characterized RF is tetherin, a protein that directly blocks the release of newly formed viral particles from infected cells. Here, we describe the evolutionary loss of a functional tetherin gene in two galliform birds, turkey ( Meleagris gallopavo ) and Mikado pheasant ( Syrmaticus mikado ). Moreover, we demonstrate that the structurally related protein TMCC(aT) exerts antiviral activity in several birds, albeit by a mechanism different from that of tetherin. The evolutionary scenario described here represents the first documented loss-of-tetherin cases in vertebrates.