RT Journal Article SR Electronic(1) A1 Burkhardt, Christiane A1 Sung, Po-Yu A1 Celma, Cristina C. A1 Roy, PollyYR 2014 T1 Structural constraints in the packaging of bluetongue virus genomic segments JF Journal of General Virology, VO 95 IS 10 SP 2240 OP 2250 DO https://doi.org/10.1099/vir.0.066647-0 PB Microbiology Society, SN 1465-2099, AB The mechanism used by bluetongue virus (BTV) to ensure the sorting and packaging of its 10 genomic segments is still poorly understood. In this study, we investigated the packaging constraints for two BTV genomic segments from two different serotypes. Segment 4 (S4) of BTV serotype 9 was mutated sequentially and packaging of mutant ssRNAs was investigated by two newly developed RNA packaging assay systems, one in vivo and the other in vitro. Modelling of the mutated ssRNA followed by biochemical data analysis suggested that a conformational motif formed by interaction of the 5′ and 3′ ends of the molecule was necessary and sufficient for packaging. A similar structural signal was also identified in S8 of BTV serotype 1. Furthermore, the same conformational analysis of secondary structures for positive-sense ssRNAs was used to generate a chimeric segment that maintained the putative packaging motif but contained unrelated internal sequences. This chimeric segment was packaged successfully, confirming that the motif identified directs the correct packaging of the segment., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.066647-0