Name
Cryo-EM Structures of Rotavirus DLPs and vDLPs Reveal Insights into Genome Packaging and Capsid Assembly
Presenter
Daniel Luque, The University of New South Wales
Co-Author(s)
Dunia Asensio-Cob1, Javier M. Rodríguez2, Daniel Luque3,4
1 Department of Molecular Medicine, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children, 686 Bay Street, Toronto ON, M5G0A4, Canada.
2 Departamento de Estructura de Macromoléculas, Centro Nacional de Biotecnología, CSIC, Cantoblanco, 28049 Madrid, Spain.
3Electron Microscope Unit, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, NSW 2052, Australia.
4 School of Biomedical Sciences, University of New South Wales, Sydney, NSW 2052, Australia.
Abstract Category
Structural Biology
Abstract
Rotavirus is a major cause of severe gastroenteritis in young children, responsible for an estimated 590 million infections annually. The structural organization of the rotavirus particle is essential for genome replication, virion assembly, and transmission. While double-layered particles (DLPs) serve as a native model of the replication machinery, genome-free double-layered virus-like particles (vDLPs) provide a valuable model system for studying capsid assembly and structure.
Here, we present the cryo-electron microscopy (cryo-EM) structures of both native DLPs and recombinant vDLPs. The high-resolution structure of DLPs reveals the ordered organization of the dsRNA genome, highlighting its structural role within the virion. Comparisons with vDLPs demonstrate strong structural conservation, validating them as a model system while also revealing subtle differences in stability and conformation due to the absence of genomic dsRNA and polymerase complexes. These differences provide mechanistic insights into RNA replication and genome packaging, supporting the hypothesis that ssRNA replication into dsRNA drives structural rearrangements.
Furthermore, intracellular purification of assembly intermediates uncovered novel structural transitions between DLPs and triple-layered particles (TLPs), shedding light on the stepwise process of particle maturation. Our findings further refine the structural framework for understanding rotavirus assembly and may help inform antiviral strategies targeting capsid morphogenesis.
Here, we present the cryo-electron microscopy (cryo-EM) structures of both native DLPs and recombinant vDLPs. The high-resolution structure of DLPs reveals the ordered organization of the dsRNA genome, highlighting its structural role within the virion. Comparisons with vDLPs demonstrate strong structural conservation, validating them as a model system while also revealing subtle differences in stability and conformation due to the absence of genomic dsRNA and polymerase complexes. These differences provide mechanistic insights into RNA replication and genome packaging, supporting the hypothesis that ssRNA replication into dsRNA drives structural rearrangements.
Furthermore, intracellular purification of assembly intermediates uncovered novel structural transitions between DLPs and triple-layered particles (TLPs), shedding light on the stepwise process of particle maturation. Our findings further refine the structural framework for understanding rotavirus assembly and may help inform antiviral strategies targeting capsid morphogenesis.