Activation of calcium/calmodulin-dependent kinase II following bovine rotavirus enterotoxin NSP۴ expression

Objective(s): The rotavirus nonstructural protein ۴ (NSP۴) is responsible for the increase in cytoplasmic calcium concentration through a phospholipase C-dependent and phospholipase C-independent pathways in infected cells. It is shown that increasing of intracellular calcium concentration in rotavirus infected cells is associated with the activation of some members of protein kinases family such as calcium/calmodulin-dependent kinase II, which plays a crucial role in replication and pathogenesis of the virus. The aim of this study was to expression bovine rotavirus NSP۴ gene in HEK۲۹۳ cell and evaluation of its biological effect related to activation of calcium/calmodulin-dependent kinase II in cell culture. Materials and Methods: MA۱۰۴ cells was used as a sensitive cell for propagation of virus and defined as a positive control. The NSP۴ gene was amplified and inserted into an expression vector, and introduced as a recombinant plasmid into HEK۲۹۳T cells. Western blot analysis was performed as a confirmation test for both expression of NSP۴ protein and activation of calcium/calmodulin-dependent kinase II. Results:Expression of NSP۴ and activated form of calcium/calmodulin-dependent kinase II were demonstrated by western blotting. Conclusion: It was shown that the expression of biologically active full- length NSP۴ protein in HEK۲۹۳T cells may be associated with some biological properties such as calcium calmodulin kinase II activation, which was indicator of rotaviruses replication and pathogenesis

کلیدواژه ها:

Calcium/calmodulin-dependent kinase II ، Expression ، Nonstructural protein ۴ (NSP۴) ، Rotavirus