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Dengue virus type 2 NS3 protease and NS2B-NS3 protease precursor induce apoptosis

Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia

Correspondence
Sazaly AbuBakar
sazaly{at}um.edu.my

Apoptosis was detected in Vero cell cultures expressing transfected dengue virus type 2 (DENV-2) genes. Approximately 17·5 and 51·5 % of cells expressing NS3 serine protease and NS2B-NS3₁₈₅ serine protease precursor protein [NS2B-NS3₁₈₅(pro)] genes, respectively, were apoptotic. The percentage of apoptotic cells was significantly higher in cell cultures expressing NS2B-NS3₁₈₅(pro). NS2B-NS3₁₈₅(pro) was detected as NS2B-NS3₁₈₅(pro)-EGFP fusion protein in cytoplasmic vesicular structures in the apoptotic cells. Site-directed mutagenesis which replaced His⁵¹ with Ala within the protease catalytic triad significantly reduced the ability of the expressed NS3 and NS2B-NS3₁₈₅(pro) to induce apoptosis. Results from the present study showed that DENV-2-encoded NS3 serine protease induces apoptosis, which is enhanced in cells expressing its precursor, NS2B-NS3₁₈₅(pro). These findings suggest the importance of NS2B as a cofactor to NS3 protease-induced apoptosis.