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| Title | Visualizing influenza A virus assembly by in situ cryo-electron tomography. |
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| Journal, issue, pages | Nat Commun, Vol. 16, Issue 1, Page 9394, Year 2025 |
| Publish date | Oct 23, 2025 |
 Authors | Moritz Wachsmuth-Melm / Sarah Peterl / Aidan O'Riain / Jana Makroczyová / Konstantin Fischer / Tim Krischuns / Sílvia Vale-Costa / Maria João Amorim / Petr Chlanda /   |
| PubMed Abstract | Influenza A virus (IAV) forms pleomorphic particles that package eight ribonucleoprotein complexes (vRNPs), each carrying a distinct RNA genome segment. vRNPs assemble in the nucleus and undergo ...Influenza A virus (IAV) forms pleomorphic particles that package eight ribonucleoprotein complexes (vRNPs), each carrying a distinct RNA genome segment. vRNPs assemble in the nucleus and undergo selective sorting during Rab11a-mediated trafficking to the plasma membrane. Virion assembly is orchestrated by matrix protein 1 (M1), which forms a layer beneath the viral envelope containing hemagglutinin (HA) and neuraminidase (NA). However, molecular details of vRNP distribution, cytosolic trafficking, and coordination of IAV assembly remains unclear. Using in situ cryo-ET, we reveal that HA-containing membranes provide Rab11a-dependent platforms for membrane-assisted vRNP clustering, reducing inter-vRNP distances. In the absence of HA, vRNPs cluster on NA-containing membranes and virus assembly remains intact, indicating that vRNP clustering and trafficking is membrane-assisted but HA independent. The characteristic 7 + 1 vRNP bundle forms concomitantly with budding and is orchestrated by M1 layer assembly that precedes plasma membrane attachment. We further reveal that intracellular M1 forms multilayered helical assemblies of antiparallel dimers, structurally distinct from the M1 layer in virions. These assemblies are compact in the nucleus but partially dissociate in the cytoplasm, likely serving as a reservoir for budding. Together, our findings uncover membrane-assisted vRNP clustering and molecular details of M1 coordinated influenza virus assembly. |
 External links | Nat Commun / PubMed:41130956 / PubMed Central |
| Methods | EM (tomography) |
| Structure data |  EMDB-50067: Tomogram showing an HA remodelled membrane in an A549wt cell infected with PR8 at 16 hpi [figure 1].  EMDB-50070: Tomogram showing an HA remodelled membrane in an A549wt cell infected with HK68 at 16 hpi [figure 1].  EMDB-50078: Tomogram showing an HA remodelled membrane in an A549wt cell infected with HK68 at 16 hpi [figure 1].  EMDB-50079: Tomogram showing an HA remodelled membrane in an A549wt cell infected with HK68 at 16 hpi [figure 1].  EMDB-50080: Tomogram showing an HA remodelled membrane in an A549wt cell infected with PR8 at 16 hpi [figure 1].  EMDB-50081: Tomogram showing an HA remodelled membrane in an A549-Rab11wt cell infected with PR8 at 16 hpi [figure 2].  EMDB-50082: A549-Rab11dn cell infected with PR8 virus at 16 hpi.  EMDB-50083: A549wt cell infected with PR8 virus at 16 hpi.  EMDB-50084: A549wt cell infected with PR8 virus at 8 hpi.  EMDB-50085: A549wt cell infected with PR8 virus at 16 hpi.  EMDB-50086: A549wt cell infected with PR8 virus at 16 hpi.  EMDB-50087: A549wt cell infected with PR8 virus at 16 hpi.  EMDB-50088: A549wt cell infected with HK68 virus at 16 hpi.  EMDB-54375: Tomogram showing an NA membrane in an A549wt cell infected with WSNdeltaHA at 16 hpi. |
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Homo sapiens (human)