CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection
ABSTRACT Ca2+/calmodulin-dependent protein kinase II (CaMKII) is one of hundreds of host-cell factors involved in the propagation of type A influenza virus (IAV), although its mechanism of action is unknown. Here, we identified CaMKII inhibitory peptide M3 by targeting its kinase domain using affini...
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American Society for Microbiology
2025-01-01
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| Series: | mBio |
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| Online Access: | https://journals.asm.org/doi/10.1128/mbio.00087-24 |
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| author | Shinichiro Hama Miho Watanabe-Takahashi Hiroki Nishimura Jumpei Omi Masakazu Tamada Takashi Saitoh Katsumi Maenaka Yuta Okuda Aoi Ikegami Asami Kitagawa Koudai Furuta Kana Izumi Eiko Shimizu Takashi Nishizono Makoto Fujiwara Tomohiro Miyasaka Shigeo Takamori Hiroshi Takayanagi Keizo Nishikawa Toshihiko Kobayashi Noriko Toyama-Sorimachi Makoto Yamashita Toshiya Senda Takatsugu Hirokawa Haruhiko Bito Kiyotaka Nishikawa |
| author_facet | Shinichiro Hama Miho Watanabe-Takahashi Hiroki Nishimura Jumpei Omi Masakazu Tamada Takashi Saitoh Katsumi Maenaka Yuta Okuda Aoi Ikegami Asami Kitagawa Koudai Furuta Kana Izumi Eiko Shimizu Takashi Nishizono Makoto Fujiwara Tomohiro Miyasaka Shigeo Takamori Hiroshi Takayanagi Keizo Nishikawa Toshihiko Kobayashi Noriko Toyama-Sorimachi Makoto Yamashita Toshiya Senda Takatsugu Hirokawa Haruhiko Bito Kiyotaka Nishikawa |
| author_sort | Shinichiro Hama |
| collection | DOAJ |
| description | ABSTRACT Ca2+/calmodulin-dependent protein kinase II (CaMKII) is one of hundreds of host-cell factors involved in the propagation of type A influenza virus (IAV), although its mechanism of action is unknown. Here, we identified CaMKII inhibitory peptide M3 by targeting its kinase domain using affinity-based screening of a tailored random peptide library. M3 inhibited IAV cytopathicity and propagation in cells by specifically inhibiting the acute-phase activation of retinoic acid-inducible gene I (RIG-I), which is uniquely regulated by CaMKII. Downstream of the RIG-I pathway activated TBK1 and then IRF3, which induced small but sufficient amounts of transcripts of the genes for IFN α/β to provide the capped 5’-ends that were used preferentially as primers to synthesize viral mRNAs by the cap-snatching mechanism. Importantly, knockout of RIG-I in cells almost completely inhibited the expression of IFN mRNAs and subsequent viral NP mRNA early in infection (up to 6 h after infection), which then protected cells from cytopathicity 24 h after infection. Thus, CaMKII-dependent acute-phase activation of RIG-I promoted IAV propagation, whereas the canonical RIG-I pathway stimulated antiviral activity by inducing large amounts of mRNA for IFNs and then for antiviral proteins later in infection. Co-administration of M3 with IAV infection rescued mice from the lethality and greatly reduced proinflammatory cytokine mRNA expression in the lung, indicating that M3 is highly effective against IAV in vivo. Thus, regulation of the CaMKII-dependent non-canonical RIG-I pathway may provide a novel host-factor-directed antiviral therapy.IMPORTANCEThe recent emergence of IAV strains resistant to commonly used therapeutic agents that target viral proteins has exacerbated the need for innovative strategies. Here, we originally identified CaMKII-inhibitory peptide M3, which efficiently inhibits IAV-lethality in vitro and in vivo. M3 specifically inhibited the acute-phase activation of RIG-I, which is a novel pathway to promote IAV propagation. Thus, this pathway acts in an opposite manner compared with the canonical RIG-I pathway, which plays essential roles in antiviral innate immune response later in infection. The CaMKII-dependent non-canonical RIG-I pathway can be a promising and novel drug target for the treatment of infections. |
| format | Article |
| id | doaj-art-d7a64009be2e4af08fd6192ac9ac992c |
| institution | Kabale University |
| issn | 2150-7511 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj-art-d7a64009be2e4af08fd6192ac9ac992c2025-01-08T14:00:38ZengAmerican Society for MicrobiologymBio2150-75112025-01-0116110.1128/mbio.00087-24CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infectionShinichiro Hama0Miho Watanabe-Takahashi1Hiroki Nishimura2Jumpei Omi3Masakazu Tamada4Takashi Saitoh5Katsumi Maenaka6Yuta Okuda7Aoi Ikegami8Asami Kitagawa9Koudai Furuta10Kana Izumi11Eiko Shimizu12Takashi Nishizono13Makoto Fujiwara14Tomohiro Miyasaka15Shigeo Takamori16Hiroshi Takayanagi17Keizo Nishikawa18Toshihiko Kobayashi19Noriko Toyama-Sorimachi20Makoto Yamashita21Toshiya Senda22Takatsugu Hirokawa23Haruhiko Bito24Kiyotaka Nishikawa25Department of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Health Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Hokkaido University of Science, Sapporo, Hokkaido, JapanLaboratory of Biomolecular Science and Center for Research and Education on Drug Discovery, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDepartment of Physiology and Anatomy, Faculty of Pharmacy, Nihon University, Funabashi, JapanLaboratory of Neural Membrane Biology, Graduate School of Brain Science, Doshisha University, Kyoto, JapanDepartment of Immunology, Graduate School of Medicine and Faculty of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, JapanDepartment of Cell Biology and Metabolic Biochemistry, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanDivision of Human Immunology, International Vaccine Design Center, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, JapanDivision of Human Immunology, International Vaccine Design Center, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, JapanDepartment of Clinical Infectious Diseases, Aichi Medical University, Nagakute, JapanStructural Biology Research Center, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki, JapanTransborder Medical Research Center, University of Tsukuba, Tsukuba, Ibaraki, JapanDepartment of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, JapanDepartment of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyotanabe, JapanABSTRACT Ca2+/calmodulin-dependent protein kinase II (CaMKII) is one of hundreds of host-cell factors involved in the propagation of type A influenza virus (IAV), although its mechanism of action is unknown. Here, we identified CaMKII inhibitory peptide M3 by targeting its kinase domain using affinity-based screening of a tailored random peptide library. M3 inhibited IAV cytopathicity and propagation in cells by specifically inhibiting the acute-phase activation of retinoic acid-inducible gene I (RIG-I), which is uniquely regulated by CaMKII. Downstream of the RIG-I pathway activated TBK1 and then IRF3, which induced small but sufficient amounts of transcripts of the genes for IFN α/β to provide the capped 5’-ends that were used preferentially as primers to synthesize viral mRNAs by the cap-snatching mechanism. Importantly, knockout of RIG-I in cells almost completely inhibited the expression of IFN mRNAs and subsequent viral NP mRNA early in infection (up to 6 h after infection), which then protected cells from cytopathicity 24 h after infection. Thus, CaMKII-dependent acute-phase activation of RIG-I promoted IAV propagation, whereas the canonical RIG-I pathway stimulated antiviral activity by inducing large amounts of mRNA for IFNs and then for antiviral proteins later in infection. Co-administration of M3 with IAV infection rescued mice from the lethality and greatly reduced proinflammatory cytokine mRNA expression in the lung, indicating that M3 is highly effective against IAV in vivo. Thus, regulation of the CaMKII-dependent non-canonical RIG-I pathway may provide a novel host-factor-directed antiviral therapy.IMPORTANCEThe recent emergence of IAV strains resistant to commonly used therapeutic agents that target viral proteins has exacerbated the need for innovative strategies. Here, we originally identified CaMKII-inhibitory peptide M3, which efficiently inhibits IAV-lethality in vitro and in vivo. M3 specifically inhibited the acute-phase activation of RIG-I, which is a novel pathway to promote IAV propagation. Thus, this pathway acts in an opposite manner compared with the canonical RIG-I pathway, which plays essential roles in antiviral innate immune response later in infection. The CaMKII-dependent non-canonical RIG-I pathway can be a promising and novel drug target for the treatment of infections.https://journals.asm.org/doi/10.1128/mbio.00087-24CaMKIIpeptide library screeninginfluenza virusRIG-Icap-snatching |
| spellingShingle | Shinichiro Hama Miho Watanabe-Takahashi Hiroki Nishimura Jumpei Omi Masakazu Tamada Takashi Saitoh Katsumi Maenaka Yuta Okuda Aoi Ikegami Asami Kitagawa Koudai Furuta Kana Izumi Eiko Shimizu Takashi Nishizono Makoto Fujiwara Tomohiro Miyasaka Shigeo Takamori Hiroshi Takayanagi Keizo Nishikawa Toshihiko Kobayashi Noriko Toyama-Sorimachi Makoto Yamashita Toshiya Senda Takatsugu Hirokawa Haruhiko Bito Kiyotaka Nishikawa CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection mBio CaMKII peptide library screening influenza virus RIG-I cap-snatching |
| title | CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection |
| title_full | CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection |
| title_fullStr | CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection |
| title_full_unstemmed | CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection |
| title_short | CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection |
| title_sort | camkii dependent non canonical rig i pathway promotes influenza virus propagation in the acute phase of infection |
| topic | CaMKII peptide library screening influenza virus RIG-I cap-snatching |
| url | https://journals.asm.org/doi/10.1128/mbio.00087-24 |
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