Abstract
Bluetongue virus core protein VP6 is an ATP hydrolysis dependent RNA helicase. However, despite much study, the precise role of VP6 within the viral capsid and its structure remain unclear. To investigate the requirement of VP6 in BTV replication, we initiated a structural and biological study. Multinuclear nuclear magnetic resonance spectra were assigned on his-tagged full-length VP6 (329 amino acid residues) as well as several truncated VP6 variants. The analysis revealed a large structured domain with two large loop regions that exhibit significant conformational exchange. One of the loops (amino acid position 34–130) could be removed without affecting the overall fold of the protein. Moreover, using a BTV reverse genetics system, it was possible to demonstrate that the VP6-truncated BTV was viable in BHK cells in the absence of any helper VP6 protein, suggesting that a large portion of this loop region is not absolutely required for BTV replication.
Highlights
Bluetongue virus (BTV), the etiological agent of Bluetongue disease of livestock, is a member of the Orbivirus genus of the Reoviridae family
In spite of the essential role of VP6 in the primary replication cycle of BTV, we show that the region between aa 34 and aa 92 of this large loop is not required for viral replication
To facilitate the nuclear magnetic resonance (NMR) study, BTV-10 VP6 was overexpressed with an N-terminal his-tag in E. coli
Summary
Bluetongue virus (BTV), the etiological agent of Bluetongue disease of livestock, is a member of the Orbivirus genus of the Reoviridae family. In addition to 7 structural proteins, BTV genome encodes 3 or 4 nonstructural proteins (NS1, NS2, NS3 and NS4) in infected host cells [1,2,3]. Structural studies have revealed their close association in a complex located at the 5-fold vertices of the VP3 subcore [9,10]. Despite considerable information regarding its enzymatic function of VP6 in vitro [11,12], little is known regarding its structure or location in the BTV core. While VP6, like VP1 and VP4, is co-purified with core from virus-infected cells, VP6 is not readily taken up into
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