Human glucose-regulated protein 78 modulates intracellular production and secretion of nonstructural protein 1 of dengue virus

Pucharee Songprakhon, Thawornchai Limjindaporn, Guey Chuen Perng, Chunya Puttikhunt, Thanawat Thaingtamtanha, Thanyaporn Dechtawewat, Sawanan Saitornuang, Chairat Uthaipibull, Sissades Thongsima, Pa Thai Yenchitsomanus, Prida Malasit, Sansanee Noisakran

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Virus-host interactions play important roles in virus infection and host cellular response. Several viruses, including dengue virus (DENV), usurp host chaperones to support their amplification and survival in the host cell. We investigated the interaction of nonstructural protein 1 (NS1) of DENV with three endoplasmic reticulum-resident chaperones (i.e. GRP78, calnexin and calreticulin) to delineate their functional roles and potential binding sites for protein complex formation. GRP78 protein showed prominent association with DENV NS1 in virus-infected Huh7 cells as evidenced by co-localization and coimmunoprecipitation assays. Further studies on the functional interaction of GRP78 protein were performed by using siRNAmediated gene knockdown in a DENV replicon transfection system. GRP78 knockdown significantly decreased intracellular NS1 production and delayed NS1 secretion but had no effect on viral RNA replication. Dissecting the important domain of GRP78 required for DENV NS1 interaction showed co-immunoprecipitation of DENV NS1 with a full-length and substratebinding domain (SBD), but not an ATPase domain, of GRP78, confirming their interaction through SBD binding. Molecular dynamics simulations of DENV NS1 and human GRP78 complex revealed their potential binding sites through hydrogen and hydrophobic bonding. The majority of GRP78-binding sites were located in a β-roll domain and connector subdomains on the DENV NS1 structure involved in hydrophobic surface formation. Taken together, our findings demonstrated the roles of human GRP78 in facilitating the intracellular production and secretion of DENV NS1 as well as predicted potential binding sites between the DENV NS1 and GRP78 complex, which could have implications in the future development of target-based antiviral drugs.

Original languageEnglish
Article number001134
Pages (from-to)1391-1406
Number of pages16
JournalJournal of General Virology
Volume99
Issue number10
DOIs
Publication statusPublished - 2018 Oct

All Science Journal Classification (ASJC) codes

  • Virology

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