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-Structure paper
タイトル | Characterization of the rotavirus assembly pathway in situ using cryoelectron tomography. |
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ジャーナル・号・ページ | Cell Host Microbe, Vol. 31, Issue 4, Page 604-615.e4, Year 2023 |
掲載日 | 2023年4月12日 |
著者 | Pranav N M Shah / James B Gilchrist / Björn O Forsberg / Alister Burt / Andrew Howe / Shyamal Mosalaganti / William Wan / Julika Radecke / Yuriy Chaban / Geoff Sutton / David I Stuart / Mark Boyce / |
PubMed 要旨 | Rotavirus assembly is a complex process that involves the stepwise acquisition of protein layers in distinct intracellular locations to form the fully assembled particle. Understanding and ...Rotavirus assembly is a complex process that involves the stepwise acquisition of protein layers in distinct intracellular locations to form the fully assembled particle. Understanding and visualization of the assembly process has been hampered by the inaccessibility of unstable intermediates. We characterize the assembly pathway of group A rotaviruses observed in situ within cryo-preserved infected cells through the use of cryoelectron tomography of cellular lamellae. Our findings demonstrate that the viral polymerase VP1 recruits viral genomes during particle assembly, as revealed by infecting with a conditionally lethal mutant. Additionally, pharmacological inhibition to arrest the transiently enveloped stage uncovered a unique conformation of the VP4 spike. Subtomogram averaging provided atomic models of four intermediate states, including a pre-packaging single-layered intermediate, the double-layered particle, the transiently enveloped double-layered particle, and the fully assembled triple-layered virus particle. In summary, these complementary approaches enable us to elucidate the discrete steps involved in forming an intracellular rotavirus particle. |
リンク | Cell Host Microbe / PubMed:36996819 / PubMed Central |
手法 | EM (単粒子) / EM (サブトモグラム平均) |
解像度 | 2.7 - 27.0 Å |
構造データ | EMDB-16146, PDB-8bp8: EMDB-16762: In situ map of Rotavirus SLP EMDB-16769: In situ STA of rotavirus DLP (penton) EMDB-16771: In situ STA of Rotavirus enveloped DLP (icos) EMDB-16772, PDB-8co6: EMDB-16773: In situ STA of rotavirus TLP (icos) EMDB-16774, PDB-8coa: |
化合物 | ChemComp-CA: ChemComp-ZN: |
由来 |
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キーワード | VIRAL PROTEIN / Rotavirus / SPA / STA |