RESUMO
Mechanisms of infection are deciphered at the host-pathogen interface.
Assuntos
Antivirais , Tratamento Farmacológico da COVID-19 , Endossomos , Neuropilina-1 , Receptores Virais , SARS-CoV-2 , Antivirais/farmacologia , Antivirais/uso terapêutico , Endossomos/virologia , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Neuropilina-1/antagonistas & inibidores , Receptores Virais/antagonistas & inibidores , SARS-CoV-2/efeitos dos fármacos , SARS-CoV-2/fisiologia , Rede trans-Golgi/virologia , Internalização do Vírus/efeitos dos fármacos , HumanosRESUMO
Endosomal sorting maintains cellular homeostasis by recycling transmembrane proteins and associated proteins and lipids (termed "cargoes") from the endosomal network to multiple subcellular destinations, including retrograde traffic to the trans-Golgi network (TGN). Viral and bacterial pathogens subvert retrograde trafficking machinery to facilitate infectivity. Here, we develop a proteomic screen to identify retrograde cargo proteins of the endosomal SNX-BAR sorting complex promoting exit 1 (ESCPE-1). Using this methodology, we identify Neuropilin-1 (NRP1), a recently characterized host factor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, as a cargo directly bound and trafficked by ESCPE-1. ESCPE-1 mediates retrograde trafficking of engineered nanoparticles functionalized with the NRP1-interacting peptide of the SARS-CoV-2 spike (S) protein. CRISPR-Cas9 deletion of ESCPE-1 subunits reduces SARS-CoV-2 infection levels in cell culture. ESCPE-1 sorting of NRP1 may therefore play a role in the intracellular membrane trafficking of NRP1-interacting viruses such as SARS-CoV-2.
Assuntos
COVID-19 , Endossomos , Interações Hospedeiro-Patógeno , Neuropilina-1 , SARS-CoV-2 , COVID-19/metabolismo , COVID-19/virologia , Sistemas CRISPR-Cas , Endossomos/virologia , Deleção de Genes , Humanos , Nanopartículas , Neuropilina-1/genética , Neuropilina-1/metabolismo , Proteômica , SARS-CoV-2/metabolismo , Nexinas de Classificação/metabolismo , Glicoproteína da Espícula de Coronavírus/metabolismoRESUMO
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), uses the viral spike (S) protein for host cell attachment and entry. The host protease furin cleaves the full-length precursor S glycoprotein into two associated polypeptides: S1 and S2. Cleavage of S generates a polybasic Arg-Arg-Ala-Arg carboxyl-terminal sequence on S1, which conforms to a C-end rule (CendR) motif that binds to cell surface neuropilin-1 (NRP1) and NRP2 receptors. We used x-ray crystallography and biochemical approaches to show that the S1 CendR motif directly bound NRP1. Blocking this interaction by RNA interference or selective inhibitors reduced SARS-CoV-2 entry and infectivity in cell culture. NRP1 thus serves as a host factor for SARS-CoV-2 infection and may potentially provide a therapeutic target for COVID-19.