Isolation and Characterization of a Novel Gammaherpesvirus from a Microbat Cell Line

Shabman, Reed S.; Shrivastava, Susmita; Tsibane, Tshidi; Attie, Oliver; Jayaprakash, Anitha; Mire, Chad E.; Dilley, Kari E.; Puri, Vinita; Stockwell, Timothy B.; Geisbert, Thomas W.; Sachidanandam, Ravi; Basler, Christopher F.

Identification and analysis of ORFs that do not share homology with EHV-2 ORFs. We sought to further characterize the features of the ORFs at the 5= end of the BGHV8 genome that share little or no homology with the EHV-2. Each ORF highlighted in blue in Fig. 4 was subjected to a BLAST search against both the viral protein database and the mammalian protein database. BLAST results against the viral protein database indicated that seven of the 10 blue ORFs were similar to proteins in other herpesviruses, and therefore, these were assigned a name standard to other gammaherpesvirus accessory genes. These include v-FLIP, E11 (v-OX2), dihydrofolate reductase (DHFR), v-Bcl-2, CCP, and MIR2 ORFs at the far 5= end of the genome (Fig. 4 ). An alignment of the BGHV8 v-OX2 aligned with the KSHV v-OX2 (Fig. 6A ) highlights conserved regions in red, and a phylogenic analysis illustrates that BGHV8 v-OX2 clusters with v-OX2 proteins of other gammaherpesviruses (Fig. 6B) . Phylogenic analysis of the other ORFs confirmed the relatedness of additional BGHV8 ORFs with other gammaherpesvirus ORFs (data not shown). In addition, a single nonhomologous ORF between E7A and ORF52 also shares similarity with select herpesvirus envelope glycoproteins. Furthermore, three ORFs in the 5= end of the genome shared no homology to any viral or mammalian protein and are therefore labeled as unknown, abbreviated U4, U5, and U6.