Inhibition of CRM1-mediated nuclear export of influenza A nucleoprotein and nuclear export protein as a novel target for antiviral drug development

• Published in: Virology (New York, N.Y.) Vol. 507; pp. 32 - 39
• Main Authors: Chutiwitoonchai, Nopporn, Mano, Takafumi, Kakisaka, Michinori, Sato, Hirotaka, Kondoh, Yasumitsu, Osada, Hiroyuki, Kotani, Osamu, Yokoyama, Masaru, Sato, Hironori, Aida, Yoko
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2017
• Subjects: 60 APPLIED LIFE SCIENCES; ABUNDANCE; Active Transport, Cell Nucleus - drug effects; antiviral agents; Antiviral Agents - pharmacology; antiviral properties; Cell Line; Cell Nucleus - metabolism; Cell Nucleus - virology; Chromosome region maintenance 1; CHROMOSOMES; computer simulation; CONCENTRATION RATIO; drug development; DRUGS; Exportin 1 Protein; FLUORESCENCE; high-throughput screening methods; Humans; IN VITRO; Infectious Disease; INFLUENZA; Influenza A virus; Influenza A virus - genetics; Influenza A virus - metabolism; INFLUENZA VIRUSES; Influenza, Human - metabolism; Influenza, Human - virology; INHIBITION; Karyopherins - antagonists & inhibitors; Karyopherins - genetics; Karyopherins - metabolism; Nuclear export protein; Nuclear export signal; Nucleoprotein; NUCLEOPROTEINS; Nucleoproteins - genetics; Nucleoproteins - metabolism; Orthomyxoviridae; protein transport; Receptors, Cytoplasmic and Nuclear - antagonists & inhibitors; Receptors, Cytoplasmic and Nuclear - genetics; Receptors, Cytoplasmic and Nuclear - metabolism; SCREENING; LMB; NP; MOE; CC50; vRNP; Nucleoprotein; CRM1; Chromosome region maintenance 1; AcGFP; Nuclear export signal; Influenza A virus; IC50; NEP; EC50; Nuclear export protein; NES; half-maximal effective concentration; IC 50; Aequorea coerulescens green fluorescent protein; chromosome region maintenance 1; nucleoprotein; nuclear export signal; viral ribonucleoprotein; Molecular Operating Environment; nuclear export protein; half-maximal inhibitory concentration; leptomycin B; CC 50; EC 50; half-maximal cytotoxic concentration
• ISSN: 0042-6822; 1096-0341; 1096-0341
• DOI: 10.1016/j.virol.2017.04.001

An anti-influenza compound, DP2392-E10 based on inhibition of the nuclear export function of the viral nucleoprotein-nuclear export signal 3 (NP-NES3) domain was successfully identified by our previous high-throughput screening system. Here, we demonstrated that DP2392-E10 exerts its antiviral effect by inhibiting replication of a broad range of influenza A subtypes. In regard to the molecular mechanism, we revealed that DP2392-E10 inhibits nuclear export of both viral NP and nuclear export protein (NEP). More specifically, in vitro pull-down assays revealed that DP2392-E10 directly binds cellular CRM1, which mediates nuclear export of NP and NEP. In silico docking suggested that DP2392-E10 binds at a region close to the HEAT9 and HEAT10 domains of CRM1. Together, these results indicate that the CRM1-mediated nuclear export function of influenza virus represents a new potential target for antiviral drug development, and also provide a core structure for a novel class of inhibitors that target this function. •DP2392-E10 inhibits replication of a broad range of influenza A subtypes.•DP2392-E10 inhibits nuclear exports of NP and NEP via their NP-NES3 and NEP-NES2 domains, respectively.•DP2392-E10 is predicted to directly bind CRM1 in the region near the HEAT9 and HEAT10 repeats.