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Structure and Sequence Requirements for RNA Capping at the Venezuelan Equine Encephalitis Virus RNA 5′ End

Abstract : Venezuelian enquine encephalitis virus (VEEV) is a reemerging arthropod-borne virus causing encephalitis in humans and domesticated animals. VEEV possesses a positive single-stranded RNA genome capped at its 5′ end. The capping process is performed by the nonstructural protein nsP1, which bears methyl and guanylyltransferase activities. The capping reaction starts with the methylation of GTP. The generated m7GTP is complexed to the enzyme to form an m7GMP-nsP1 covalent intermediate. The m7GMP is then transferred onto the 5′-diphosphate end of the viral RNA. Here, we explore the specificities of the acceptor substrate in terms of length, RNA secondary structure, and/or sequence. Any diphosphate nucleosides but GDP can serve as acceptors of the m7GMP to yield m7GpppA, m7GpppC, or m7GpppU. We show that capping is more efficient on small RNA molecules, whereas RNAs longer than 130 nucleotides are barely capped by the enzyme. The structure and sequence of the short, conserved stem-loop, downstream to the cap, is an essential regulatory element for the capping process.
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https://hal-amu.archives-ouvertes.fr/hal-03426112
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Submitted on : Friday, November 12, 2021 - 8:34:53 AM
Last modification on : Tuesday, November 16, 2021 - 8:35:44 AM

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Oney Ortega Granda, Coralie Valle, Ashleigh Shannon, Etienne Decroly, Bruno Canard, et al.. Structure and Sequence Requirements for RNA Capping at the Venezuelan Equine Encephalitis Virus RNA 5′ End. Journal of Virology, American Society for Microbiology, 2021, 95 (15), ⟨10.1128/JVI.00777-21⟩. ⟨hal-03426112⟩

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