Structural basis and dynamics of Chikungunya alphavirus RNA capping by nsP1 capping pores - Archive ouverte HAL Access content directly
Journal Articles Proceedings of the National Academy of Sciences of the United States of America Year : 2023

Structural basis and dynamics of Chikungunya alphavirus RNA capping by nsP1 capping pores

Rhian Jones
  • Function : Author
Michael Hons
  • Function : Author
Nadia Rabah
Noelia Zamarreño
Rocío Arranz
Juan Reguera

Abstract

Alphaviruses are emerging positive-stranded RNA viruses which replicate and transcribe their genomes in membranous organelles formed in the cell cytoplasm. The nonstructural protein 1 (nsP1) is responsible for viral RNA capping and gates the replication organelles by assembling into monotopic membrane-associated dodecameric pores. The capping pathway is unique to Alphaviruses; beginning with the N 7 methylation of a guanosine triphosphate (GTP) molecule, followed by the covalent linkage of an m 7 GMP group to a conserved histidine in nsP1 and the transfer of this cap structure to a diphosphate RNA. Here, we provide structural snapshots of different stages of the reaction pathway showing how nsP1 pores recognize the substrates of the methyl-transfer reaction, GTP and S-adenosyl methionine (SAM), how the enzyme reaches a metastable postmethylation state with SAH and m 7 GTP in the active site, and the subsequent covalent transfer of m 7 GMP to nsP1 triggered by the presence of RNA and postdecapping reaction conformational changes inducing the opening of the pore. In addition, we biochemically characterize the capping reaction, demonstrating specificity for the RNA substrate and the reversibility of the cap transfer resulting in decapping activity and the release of reaction intermediates. Our data identify the molecular determinants allowing each pathway transition, providing an explanation for the need for the SAM methyl donor all along the pathway and clues about the conformational rearrangements associated to the enzymatic activity of nsP1. Together, our results set ground for the structural and functional understanding of alphavirus RNA-capping and the design of antivirals.

Dates and versions

hal-04061869 , version 1 (07-04-2023)

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Rhian Jones, Michael Hons, Nadia Rabah, Noelia Zamarreño, Rocío Arranz, et al.. Structural basis and dynamics of Chikungunya alphavirus RNA capping by nsP1 capping pores. Proceedings of the National Academy of Sciences of the United States of America, 2023, 120 (12), ⟨10.1073/pnas.2213934120⟩. ⟨hal-04061869⟩

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