Mechanism Of AP-2-associated Protein Kinase 1 Regulates Rabies Virus Infection

Rabies is caused by rabies virus(RABV)infection and is a neglected but important zoonotic disease.Once the clinical symptoms of RABV infection appear,the mortality rate is almost 100%.Rabies kills 59,000 people worldwide each year.Rabies can be effectively prevented by vaccines and immunoglobulins,but there is no effective drug to treat patients with clinical symptoms currently.A global plan for rabies control and elimination was agreed by FAO,the OIE and WHO:The Global Strategic Plan to End Human Deaths from Dog-mediated Rabies by 2030.Carrying out research on the pathogenic mechanism of RABV,in-depth exploration of important factors in the process of RABV infection,and finding potential drug targets,have important reference value for the treatment of rabies.We carried out a high-through-put(HPT)RNAi assay in attempt to identify cellular factors that affects RABV entry into the cells and identified AP2-associated kinase 1(AAK1),a serine/threonine kinase,as an important cellular component in regulating the entry of RABV.AAK1 knock-down greatly inhibits RABV infection in human embryonic kidney cells(HEK293),human neuroblastoma cells(SK-N-SH),and mouse cerebral cortex primary neurons(mPN).RABV invades the cell through clathrin-mediated endocytosis(CME).AAK1 is host kinase that regulate phosphorylation of the clathrin-associated adaptor complex AP2 ? subunit(AP2M1).Thus,we hypothesize that AAK1 regulate RABV infection dependently of its involvement in clathrin-mediated endocytosis.In this study,we first confirm the suppression of RABV infection in AAK1-and AP2M1-knockdown cells.We observed the co-localization of RABV with AP2 or Rab5 in cells treated or untreated with AAK1 inhibitor,combined with the addition of AAK1 inhibitor,it is determined that AAK1 affects the very early stage of the RABV infection.Finally,we verified that the AAK1 inhibitor sunitinib can prolong the survival but not prevents the death of mice challenged with RABV street virus.This study reveals the mechanism of AAK1 affecting RABV infection,and provides a potential drug target for the treatment of rabies.