The Rac1-PAK1-Arp2/3 signaling axis regulates CHIKV nsP1-induced filopodia and optimal viral genome replication.

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  • Additional Information
    • Source:
      Publisher: American Society For Microbiology Country of Publication: United States NLM ID: 0113724 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-5514 (Electronic) Linking ISSN: 0022538X NLM ISO Abbreviation: J Virol Subsets: MEDLINE
    • Publication Information:
      Publication: Washington Dc : American Society For Microbiology
      Original Publication: Baltimore, American Society for Microbiology.
    • Subject Terms:
      Virus Replication* ; rac1 GTP-Binding Protein*/metabolism ; rac1 GTP-Binding Protein*/genetics ; p21-Activated Kinases*/metabolism ; p21-Activated Kinases*/genetics ; Chikungunya virus*/physiology ; Chikungunya virus*/genetics ; Signal Transduction* ; Actin-Related Protein 2-3 Complex*/metabolism ; Actin-Related Protein 2-3 Complex*/genetics ; Pseudopodia*/metabolism ; Pseudopodia*/virology; Humans ; Animals ; Viral Nonstructural Proteins/metabolism ; Viral Nonstructural Proteins/genetics ; Genome, Viral ; Cell Membrane/metabolism ; Cell Membrane/virology ; Cell Line ; Chlorocebus aethiops ; Host-Pathogen Interactions ; Actin-Related Protein 2/metabolism ; Actin-Related Protein 2/genetics
    • Abstract:
      Alphavirus infection induces dramatic remodeling of host cellular membranes, producing filopodia-like and intercellular extensions. The formation of filopodia-like extensions has been primarily assigned to the replication protein nsP1, which binds and reshapes the host plasma membrane when expressed alone. While reported decades ago, the molecular mechanisms behind nsP1 membrane deformation remain unknown. Using mammalian epithelial cells and Chikungunya virus (CHIKV) as models, we characterized nsP1-induced membrane deformations as highly dynamic actin-rich lamellipodia and filopodia-like extensions. Through pharmacological inhibition and genetic invalidation, we identified the critical contribution of the Rac1 GTPase and its downstream effectors PAK1 and the actin nucleator Arp2 in nsP1-induced membrane deformation. An intact Rac1-PAK1-Arp2 signaling axis was also required for optimal CHIKV genome replication. Therefore, our results designate the Rac1-PAK1-Arp2 pathway as an essential signaling node for CHIKV infection and establish a parallel requirement for host factors involved in nsP1-induced plasma membrane reshaping and assembly of a functional replication complex.IMPORTANCEThe alphavirus nsP1 protein dramatically remodels host cellular membranes, resulting in the formation of filopodia-like extensions. Although described decades ago, the molecular mechanisms controlling these membrane deformations and their functional importance remain elusive. Our study provides mechanistic insight, uncovering the critical role of the Rac1 GTPase, along with its downstream effectors PAK1 and the actin nucleator Arp2, in the nsP1-associated phenotype. Furthermore, we demonstrate that the Rac1-PAK1-Arp2 pathway is essential for optimal CHIKV genome replication. Our findings establish a parallel in the cellular mechanisms governing nsP1-induced plasma membrane reshaping and the production of a functional replication complex in infected cells.
      Competing Interests: The authors declare no conflict of interest.
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    • Grant Information:
      ANR-18-CE11-0026-01 Agence Nationale de la Recherche (ANR); Fondation pour la Recherche Médicale (FRM); Mediterranee Infection, Infectiopole Sud Foundation
    • Contributed Indexing:
      Keywords: Chikungunya virus; alphavirus; cell signaling; cytoskeleton; genome replication; membrane rearrangement; nonstructural protein 1; nsP1
    • Accession Number:
      EC 3.6.5.2 (rac1 GTP-Binding Protein)
      EC 2.7.11.1 (p21-Activated Kinases)
      0 (RAC1 protein, human)
      EC 2.7.11.1 (PAK1 protein, human)
      0 (Actin-Related Protein 2-3 Complex)
      0 (Viral Nonstructural Proteins)
      0 (Actin-Related Protein 2)
    • Publication Date:
      Date Created: 20240919 Date Completed: 20241022 Latest Revision: 20241024
    • Publication Date:
      20241025
    • Accession Number:
      PMC11495065
    • Accession Number:
      10.1128/jvi.00612-24
    • Accession Number:
      39297643