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Home / Equine Research Bank / J Virol / Structure and Sequence Requirements for RNA Capping at the V...
Journal of virology2021; 95(15); e0077721; doi: 10.1128/JVI.00777-21

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

Abstract: Venezuelan equine 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 mGTP is complexed to the enzyme to form an mGMP-nsP1 covalent intermediate. The mGMP 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 mGMP to yield mGpppA, mGpppC, or mGpppU. 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. The emergence, reemergence, and expansion of alphaviruses (genus of the family ) are a serious public health and epizootic threat. Venezuelan equine encephalitis virus (VEEV) causes encephalitis in human and domesticated animals, with a mortality rate reaching 80% in horses. To date, no efficient vaccine or safe antivirals are available for human use. VEEV nonstructural protein 1 (nsP1) is the viral capping enzyme characteristic of the genus. nsP1 catalyzes methyltransferase and guanylyltransferase reactions, representing a good therapeutic target. In the present report, we provide insights into the molecular features and specificities of the cap acceptor substrate for the guanylylation reaction.
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Publication Date: 2021-07-12 PubMed ID: 34011549PubMed Central: PMC8274618DOI: 10.1128/JVI.00777-21Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research explores the specific characteristics of Venezuelan Equine Encephalitis Virus (VEEV)’s RNA capping process, performed by its nonstructural protein nsP1. This study intends to offer insights regarding the RNA’s length, sequence, and structure necessities for this process.

Objective of the Research

  • The primary purpose of this research is to understand the specific requirements and properties of the Venezuelan Equine Encephalitis Virus (VEEV)’s RNA capping process. This process is instrumental in the virus’s ability to cause encephalitis in humans and animals.

Procedure and Findings

  • The capping process is carried out by a nonstructural protein referred to as nsP1. This protein exhibits guanylyltransferase and methyltransferase activities – two crucial reactions in the virus’s replication process.
  • The capping reaction begins with GTP’s methylation. The newly generated mGTP then connects to the enzyme to form an mGMP-nsP1 covalent intermediate. Finally, the mGMP is transferred onto the viral RNA’s 5′-diphosphate end.
  • The researchers explore the specificities of the substrate that accepts this mGMP in the capping process, particularly concerning its length, sequence, and secondary structure.
  • The study shows that the capping process is more effective on smaller RNA molecules. In contrast, RNAs that are longer than 130 nucleotides are barely capped by the enzyme.
  • They also discover that the conserved stem-loop, located downstream to the cap, is critical for the capping process. Its structure and sequence serve as a vital regulatory element.

Significance of the Research

  • This research offers valuable insight into the mechanisms that enable the VEEV to cause severe diseases in humans and animals. The information can be instrumental in the development of effective vaccines and antivirals.
  • Alphaviruses are a serious public health and epizootic threat due to their ability to emerge, reemerge, and expand. VEEV, a type of Alphavirus, has a high mortality rate, with little to no efficient vaccine or safe antiviral available for human use.
  • The VEEV’s nonstructural protein nsP1, which plays a pivotal role in the virus’s propagation, could potentially serve as an excellent therapeutic target. The research provides a foundation for understanding the molecular features of the cap acceptor substrate, critical for select therapeutic interventions targeting the guanylylation reaction.

Cite This Article

APA
Ortega Granda O, Valle C, Shannon A, Decroly E, Canard B, Coutard B, Rabah N. (2021). Structure and Sequence Requirements for RNA Capping at the Venezuelan Equine Encephalitis Virus RNA 5′ End. J Virol, 95(15), e0077721. https://doi.org/10.1128/JVI.00777-21

Publication

Journal of virology
ISSN: 1098-5514
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 95
Issue: 15
Pages: e0077721
PII: e00777-21

Researcher Affiliations

Ortega Granda, Oney
  • Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France.
Valle, Coralie
  • Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France.
Shannon, Ashleigh
  • Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France.
Decroly, Etienne
  • Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France.
Canard, Bruno
  • Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France.
Coutard, Bruno
  • Unité des Virus Emergents (UVE): Aix-Marseille Univ-IRD 190-Inserm, Marseille, France.
Rabah, Nadia
  • Aix Marseille Université, CNRS, AFMB UMR 7257, Marseille, France.
  • Université de Toulon, La Garde, France.

MeSH Terms

  • Animals
  • Encephalitis Virus, Venezuelan Equine / genetics
  • Encephalomyelitis, Venezuelan Equine / pathology
  • Encephalomyelitis, Venezuelan Equine / virology
  • Horses
  • Humans
  • Methyltransferases / metabolism
  • Nucleic Acid Conformation
  • Nucleotidyltransferases / metabolism
  • RNA Caps / genetics
  • RNA, Viral / genetics
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism
  • Virus Replication / genetics

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Citations

This article has been cited 1 times.
  1. Jones R, Hons M, Rabah N, Zamarreño N, Arranz R, Reguera J. Structural basis and dynamics of Chikungunya alphavirus RNA capping by nsP1 capping pores.. Proc Natl Acad Sci U S A 2023 Mar 21;120(12):e2213934120.
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