Study of Hsp90α and Hsp90β role in virus replication using cell lines with Hsp90 gene knockouts

Replication of the human Enterovirus 71 (EV71) and herpes simplex virus 1 (HSV-1) requires Hsp90 chaperone activity. Vertebrate cells express two cytosolic Hsp90 proteins, Hsp90α and Hsp90β. Earlier reports suggested that EV71 replication might depend solely on the Hsp90β, whereas HSV-1 replication depended on Hsp90α. Here, we describe construction of the cell line knockouts missing Hsp90α or Hsp90β protein. Using these cells, we found that HSV-1 and, another enterovirus, Coxsackievirus B5 (CVB5) replicate in both Hsp90α and Hsp90β knockout cells with equal efficiency. The presented results demonstrate that cell lines with a mutation inactivating the specific HSP90 gene might be an easy-to-use and robust system to study specific cellular functions of Hsp90α and Hsp90β.