2018 Fiscal Year Research-status Report
Studies on influenza A virus M2-host interactome and its role in virus replication
| Project/Area Number |
16K08014
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| Research Institution | Hokkaido University |
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Principal Investigator |
マンズール ラシッド 北海道大学, 人獣共通感染症リサーチセンター, 特任助教 (90566150)
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| Project Period (FY) |
2016-04-01 – 2020-03-31
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| Keywords | Influenza A virus / M2 / Host interaction |
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| Outline of Annual Research Achievements |
Influenza A virus (IAV) matrix protein 2 (M2) is an integral membrane, ion channel protein. IAVs express three viral envelop proteins in infected cells i.e. hemagglutinin (HA) and neuraminidase (NA) & M2. Though all three viral envelop proteins are abundantly expressed in infected cells, only HA and NA become the major viral envelop proteins. Both HA and NA are targeted to the lipid rafts, whereas M2 is excluded from lipid rafts. The transmembrane domains (TMDs) of M2, HA and NA are 19, 27 and 29 residues long. Despite the presence of two host-cell membrane lipid-raft targeting features i.e., palmitoylation at Cys50 and cholesterol recognition/interaction amino acid consensus (CRAC) motif, relatively shorter M2-TMD is believed to prevent its association with lipid-raft domains. Therefore, we investigated the mechanism of interaction/incorporation of M2-TMD in non-raft regions of host cell membrane.
So far, we have generated M2-TMD mutants by inserting 3, 6 and 8 amino acid (aa) residues in N-terminal end of TMD. We also introduced similar mutations in M gene for generating viruses by reverse genetics. Then, we characterized the M2-TMD mutants by comparing the cell surface expression, cytotoxic potential and lipid raft association of tested M2 mutants with that of wtM2 protein. It was found that M2 mutants , like wtM2, were expressed on cell surface and displayed cytotoxic potential. Interestingly, it was found that increasing the TMD length did not affect their non-lipid raft association character, and like wtM2, M2-TMD mutants remained in the non-raft microdomains.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
Additional assay condition is being optimized to assess the interaction of M2-TMD mutants with host cell membrane microdomain environment. Attempts to rescue the viruses by reverse genetics being are made. Manuscript is also being prepared.
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| Strategy for Future Research Activity |
1. To examine the effect of increase in the length of M2 transmembrane domain by insertion mutations on virus release by techniques such as real-time PCR, electron microscopy, etc.
2. To examine the effectof increase in the length of M2 transmembrane domain by insertion mutations on virus replication kinetics.
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| Causes of Carryover |
Slight change in research plan to understand the interaction of M2 transmembrane domain, in addition to M2-cytoplasmic tail and ectodomain, with host cellular factors.
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Research Products
(3 results)
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[Journal Article] Generation of bat-derived influenza viruses and their reassortants2019
Author(s)
Masahiro Sato, Junki Maruyama, Tatsunari Kondoh, Naganori Nao, Hiroko Miyamoto, Yoshihiro Takadate, Wakako Furuyama, Masahiro Kajihara, Hirohito Ogawa, Rashid Manzoor, Reiko Yoshida, Manabu Igarashi and Ayato Takada
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Journal Title
Scientific Reports
Volume: 9
Pages: 1-8
DOI
Peer Reviewed / Open Access / Int'l Joint Research
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