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Home / Equine Research Bank / J Virol / Efficient homologous RNA recombination and requirement for a...
Journal of virology2000; 74(19); 9062-9070; doi: 10.1128/jvi.74.19.9062-9070.2000

Efficient homologous RNA recombination and requirement for an open reading frame during replication of equine arteritis virus defective interfering RNAs.

Abstract: Equine arteritis virus (EAV), the prototype arterivirus, is an enveloped plus-strand RNA virus with a genome of approximately 13 kb. Based on similarities in genome organization and protein expression, the arteriviruses have recently been grouped together with the coronaviruses and toroviruses in the newly established order Nidovirales. Previously, we reported the construction of pEDI, a full-length cDNA copy of EAV DI-b, a natural defective interfering (DI) RNA of 5.6 kb (R. Molenkamp et al., J. Virol. 74:3156-3165, 2000). EDI RNA consists of three noncontiguous parts of the EAV genome fused in frame with respect to the replicase gene. As a result, EDI RNA contains a truncated replicase open reading frame (EDI-ORF) and encodes a truncated replicase polyprotein. Since some coronavirus DI RNAs require the presence of an ORF for their efficient propagation, we have analyzed the importance of the EDI-ORF in EDI RNA replication. The EDI-ORF was disrupted at different positions by the introduction of frameshift mutations. These were found either to block DI RNA replication completely or to be removed within one virus passage, probably due to homologous recombination with the helper virus genome. Using recombination assays based on EDI RNA and full-length EAV genomes containing specific mutations, the rates of homologous RNA recombination in the 3'- and 5'-proximal regions of the EAV genome were studied. Remarkably, the recombination frequency in the 5'-proximal region was found to be approximately 100-fold lower than that in the 3'-proximal part of the genome.
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Publication Date: 2000-09-12 PubMed ID: 10982351PubMed Central: PMC102103DOI: 10.1128/jvi.74.19.9062-9070.2000Google Scholar: Lookup
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  • Non-U.S. Gov't

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.

This research explores the replication of defective interfering RNAs (DI RNAs) within the Equine Arteritis Virus (EAV). Among its significant findings is the necessary presence of an open reading frame (ORF) for effective RNA replication and the observation that homologous RNA recombination occurs at varying rates in different regions of the EAV genome.

Study of Equine Arteritis Virus

  • The research focuses on the Equine Arteritis Virus (EAV), a plus-strand RNA virus with a genome approximately 13 kilobases in length.
  • Arteriviruses, such as EAV, have recently been included with coronaviruses and toroviruses in the Nidovirales order due to parallels in genome organization and protein expression.

Role of Defective Interfering RNAs

  • This study examines defective interfering RNAs (DI RNAs), specifically EDI, a natural DI RNA of EAV.
  • DI RNAs are smaller viral RNAs that disrupt the replication of the parent virus. In the case of EAV, the DI RNA (EDI) includes sections of the EAV genome and contains a truncated version of the replicase gene, contributing to a reduced replicase polyprotein.

Significance of Open Reading Frame

  • The researchers tested the importance of an open reading frame (ORF) in the EDI RNA replication process.
  • A series of frameshift mutations introduced at different positions within the EDI-ORF caused either complete blocking of the DI RNA replication or removal within one virus passage via homologous recombination with the helper virus genome.

Differential Rates of Homologous Recombination

  • The study also investigates homologous RNA recombination, a process where identical or nearly identical genetic sequences exchange genetic material.
  • The researchers used recombination assays based on EDI RNA and full-length EAV genomes with specific mutations to analyze the recombination rate.
  • The findings reveal a striking 100-fold difference in recombination frequency between the 3′- and 5′-proximal regions, with the 5′-proximal region showing significantly lower recombination frequency.

Cite This Article

APA
Molenkamp R, Greve S, Spaan WJ, Snijder EJ. (2000). Efficient homologous RNA recombination and requirement for an open reading frame during replication of equine arteritis virus defective interfering RNAs. J Virol, 74(19), 9062-9070. https://doi.org/10.1128/jvi.74.19.9062-9070.2000

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 74
Issue: 19
Pages: 9062-9070

Researcher Affiliations

Molenkamp, R
  • Department of Virology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
Greve, S
    Spaan, W J
      Snijder, E J

        MeSH Terms

        • Animals
        • Equartevirus / physiology
        • Gene Expression Regulation, Viral
        • Open Reading Frames / genetics
        • RNA, Viral / genetics
        • Recombination, Genetic
        • Virus Replication

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