Probing the Sites of Interactions of Rotaviral Proteins Involved in Replication

Viskovska M, et al., 88(21):12866-81, J Virol, 2014

This investigation reveals additional protein partners that are involved and directly interact with the non-structural protein 2 (NSP2) during the replication and packaging of rotavirus genome in cytoplasmic compartments called viroplasms. These viroplasms generally formed during virus infection. A complex interactome coordinates the replication and packaging processes by making use of the non-structural proteins (NSPs) and structural proteins (VPs). With the help of isothermal calorimetry (ITC), biolayer interferometry (BLI), and peptide array screening, they examined the interactions between NSP2, NSP5, NSP6, VP1, and VP2. The BLI experiments carried out using an Octet RED96 instrument utilized biotinylated proteins immobilized on to SA biosensors. The binding assays carried out between NSP2-VP1, NSP2-VP2, and NSP2-NSP6 involved immobilization of VP1, NSP2, and NSP6, respectively on to SA sensors.

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