Lipid rafts play an important role in the early stage of severe acute respiratory syndrome-coronavirus life cycle.
Lipid rafts are involved in the life cycle of many viruses. In this study, we showed that lipid rafts also play an important role in the life cycle of severe acute respiratory syndrome (SARS)-coronavirus (CoV). Cholesterol depletion by pretreatment of Vero E6 cells with methyl-beta-cyclodextrin (MbetaCD) inhibited the production of SARS-CoV particles released from the infected cells. This inhibition was prevented by addition of cholesterol to the culture medium, indicating that the reduction of virus particle release was caused by the loss of cholesterol in the cell membrane. In contrast, cholesterol depletion at the post-entry stage (3h post-infection) caused only a limited effect on virus particle release. Northern blot analysis revealed that the levels of viral mRNAs were significantly affected by pretreatment with MbetaCD, but not by treatment at 3h post-infection. Interestingly, no apparent evidence for colocalization of angiotensin converting enzyme 2 with lipid rafts in the membrane of Vero E6 cells was obtained. These results suggest that lipid rafts could contribute to SARS-CoV infection in the early replication process in Vero E6 cells.
Li
Gui-Mei
GM
Department of Virology, Research Institute for Microbial Diseases (RIMD), Osaka University, Suita, Osaka 565-0871, Japan.
Li
Yong-Gang
YG
Yamate
Masanobu
M
Li
Shu-Ming
SM
Ikuta
Kazuyoshi
K
D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D002522:Chlorocebus aethiops; D002784:Cholesterol / Q000172:deficiency / Q000378:metabolism / Q000494:pharmacology; D005434:Flow Cytometry; D008018:Life Cycle Stages; D021962:Membrane Microdomains / Q000378:metabolism / Q000821:virology*; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D045169:Severe Acute Respiratory Syndrome / Q000378:metabolism / Q000821:virology; D014709:Vero Cells; D014779:Virus Replication / Q000187:drug effects / Q000502:physiology; D047392:beta-Cyclodextrins / Q000494:pharmacology
D012333:RNA, Messenger; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D002784:Cholesterol; D007703:Peptidyl-Dipeptidase A; D000085962:Angiotensin-Converting Enzyme 2