Structure of the N-RNA/P interface reveals mode of L/P attachment to the nucleocapsid of human metapneumovirus - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement
Preprints, Working Papers, ... Year : 2023

Structure of the N-RNA/P interface reveals mode of L/P attachment to the nucleocapsid of human metapneumovirus

Abstract

Human metapneumovirus (HMPV) is a major cause of respiratory illness in young children.

The polymerase complex of HMPV consists of two obligate components, the L polymerase and its cofactor, the phosphoprotein P. During replication and transcription, the L/P complex traverses the viral RNA genome, which is encapsidated within multimerized N nucleoproteins.

An essential interaction between N and a C-terminal region of P is required for tethering of the L/P polymerase to the RNA template. This N-P interaction is also involved in the formation of cytoplasmic viral factories in infected cells, called inclusion bodies. To define how L/P recognizes N-encapsidated RNA (N-RNA) we employed cryogenic electron microscopy (cryo-EM) and molecular dynamics simulations, coupled to polymerase activity assays and imaging of inclusion bodies in transfected cells. We report a 2.9 Å resolution structure of a triplecomplex between multimeric N, bound to both RNA and the C-terminal region of P. Furthermore, we also present cryo-EM structures of assembled N in different oligomeric states, highlighting the plasticity of N. Combined with our functional assays, these structural data delineate in molecular detail how P attaches to N-RNA whilst retaining substantial conformational dynamics. Moreover, the N-RNA-P triple complex structure provides a molecular blueprint for the design of therapeutics to potentially disrupt the attachment of L/P to its template.

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hal-04689372 , version 1 (05-09-2024)

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Jack D Whitehead, Hortense Decool, Cédric Leyrat, Loic Carrique, Jenna Fix, et al.. Structure of the N-RNA/P interface reveals mode of L/P attachment to the nucleocapsid of human metapneumovirus. 2024. ⟨hal-04689372⟩
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