Analysis of the effect of Sphingomyelinase on rubellavirus infectivity in two cell lines

Noriyuki Otsuki; Masafumi Sakata; Yoshio Mori; Kiyoko Okamoto; Makoto Takeda

doi:10.21769/BioProtoc.2992

Analysis of the effect of Sphingomyelinase on rubellavirus infectivity in two cell lines

Bio Protoc. 2018 Sep 5;8(17):e2992. doi: 10.21769/BioProtoc.2992.

Authors

Noriyuki Otsuki¹, Masafumi Sakata¹, Yoshio Mori¹, Kiyoko Okamoto¹, Makoto Takeda¹

Affiliation

¹ Department of Virology III, National Institute of Infectious Diseases, Tokyo, Japan.

Abstract

Rubella is a mildly contagious disease characterized by low-grade fever and a morbilliform rash caused by the rubella virus (RuV). Viruses often use cellular phospholipids for infection. We studied the roles of cellular sphingomyelin in RuV infection. Treatment of cells with sphingomyelinase (SMase) inhibited RuV infection in rabbit kidney-derived RK13 cells and African green monkey (Cercopithecus aethiops) kidney-derived Vero cells. Our data further demonstrated that RuV used cellular sphingomyelin and cholesterol for its binding to cells and membrane fusion at the step of virus entry. Detailed protocols of our assays, which assess the effects of SMase treatment on RuV infectivity in RK13 and Vero cells, are described.

Keywords: Lipid; Rubella virus; Sphingomyelin; Sphingomyelinase; Virus entry.