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Equine arteritis virus subgenomic RNA transcription: UV inactivation and translation inhibition studies.
Abstract: The expression of the genetic information of equine arteritis virus (EAV), an arterivirus, involves the synthesis of six subgenomic (sg) mRNAs. These are 5' and 3' coterminal since they are composed of a leader and a body sequence, which are identical to the 5' and 3' ends of the genome, respectively. Previously, it has been suggested that cis-splicing of a genome-length precursor RNA is involved in their synthesis. This was reevaluated in a comparative analysis of the sg RNA synthesis of EAV, the coronavirus mouse hepatitis virus (MHV), and the alphavirus Sindbis virus. UV transcription mapping showed that the majority of the EAV sg RNAs made at later stages of infection is not derived from a genome-length precursor. However, complete independence of sg RNA synthesis from that of genomic RNA was never observed during the course of infection. The possibility that this resulted from UV irradiation-induced effects on the synthesis of the viral replicase was investigated by inhibiting translation using cycloheximide. For EAV, ongoing protein synthesis was found to be more important for the synthesis of sg RNA than for that of genomic RNA. In general, MHV transcription was extremely sensitive to translation inhibition, whereas EAV genomic RNA synthesis became independent of de novo protein synthesis late in infection.
Publication Date: 1995-11-10 PubMed ID: 7491761PubMed Central: PMC7131190DOI: 10.1006/viro.1995.0009Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The study explores the expression of genetic information in equine arteritis virus (EAV), particularly focusing on the creation of six subgenomic mRNAs. Using UV transcription mapping and translation inhibition methods, the researchers discovered that the majority of EAV’s sg RNAs are not derived from a genome-length precursor. They also found that ongoing protein synthesis is more essential for the synthesis of sg RNA than genomic RNA.
Explanation of the Research
- This research primarily focuses on the genetic expression of the equine arteritis virus (EAV), which is part of the arterivirus family. The process of its genetic expression involves synthesizing six subgenomic, or sg, mRNAs, which are parts of the RNA composed of a lead and a body sequence. These sequences are identical to the 5′ and 3′ ends of the genome, indicating that both ends of the sequences are the same.
- Early theories suggested that the synthesis may involve cis-splicing of a genome-length precursor RNA. Cis-splicing is a method of RNA processing, which permits one single gene to code for multiple proteins in different cellular situations. This study sought to reassess these suggestions and conducted a comparative analysis using the coronavirus mouse hepatitis virus (MHV), and the alphavirus Sindbis virus.
- UV transcription mapping, a method that uses UV radiation to control gene expression, was implemented for this investigation. Through this process, they noted that the majority of EAV’s sg RNAs made at later stages of the infection did not originate from a genome-length precursor. However, they found that total independence of sg RNA synthesis from the genomic RNA synthesis was never observed during infection.
- The researchers decided to probe further into the effect of UV radiation on the synthesis of the viral replicase, the enzyme that aids in viral replication. This involved application of cycloheximide, an inhibitor of translation in eukaryotic organisms. The results suggested that ongoing protein synthesis was more crucial to the synthesis of sg RNA than the synthesis of genomic RNA for EAV.
- Cycloheximide affected EAV genomic RNA synthesis as it appears to be more dependent on de novo protein synthesis in early infection stages. Meanwhile, MHV’s transcription appeared to be extremely sensitive to translation inhibition. However, over time, EAV’s genomic RNA synthesis became less reliant on de novo protein synthesis.
Cite This Article
APA
Den Boon JA, Spaan WJ, Snijder EJ.
(1995).
Equine arteritis virus subgenomic RNA transcription: UV inactivation and translation inhibition studies.
Virology, 213(2), 364-372.
https://doi.org/10.1006/viro.1995.0009 Publication
Researcher Affiliations
- Department of Virology, Faculty of Medicine, Leiden University, The Netherlands.
MeSH Terms
- Animals
- Cell Line
- Cycloheximide / pharmacology
- Equartevirus / genetics
- Equartevirus / physiology
- Equartevirus / radiation effects
- Genome, Viral
- Mice
- Murine hepatitis virus / genetics
- Murine hepatitis virus / radiation effects
- Protein Biosynthesis
- Protein Synthesis Inhibitors / pharmacology
- RNA Precursors / metabolism
- RNA, Messenger / biosynthesis
- RNA, Viral / biosynthesis
- RNA, Viral / genetics
- RNA, Viral / radiation effects
- RNA-Dependent RNA Polymerase / biosynthesis
- Sindbis Virus / genetics
- Sindbis Virus / radiation effects
- Transcription, Genetic
- Ultraviolet Rays
- Viral Proteins / biosynthesis
Citations
This article has been cited 14 times.- Kuiper JWP, Baade T, Kremer M, Kranaster R, Irmisch L, Schuchmann M, Zander J, Marx A, Hauck CR. Detection of SARS-CoV-2 from raw patient samples by coupled high temperature reverse transcription and amplification. PLoS One 2020;15(11):e0241740.
- Bello-Onaghise G, Wang G, Han X, Nsabimana E, Cui W, Yu F, Zhang Y, Wang L, Li Z, Cai X, Li Y. Antiviral Strategies of Chinese Herbal Medicine Against PRRSV Infection. Front Microbiol 2020;11:1756.
- Nedialkova DD, Gorbalenya AE, Snijder EJ. Arterivirus Nsp1 modulates the accumulation of minus-strand templates to control the relative abundance of viral mRNAs. PLoS Pathog 2010 Feb 19;6(2):e1000772.
- Sawicki SG, Sawicki DL. Coronavirus transcription: a perspective. Curr Top Microbiol Immunol 2005;287:31-55.
- van Dinten LC, van Tol H, Gorbalenya AE, Snijder EJ. The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis. J Virol 2000 Jun;74(11):5213-23.
- Molenkamp R, Rozier BC, Greve S, Spaan WJ, Snijder EJ. Isolation and characterization of an arterivirus defective interfering RNA genome. J Virol 2000 Apr;74(7):3156-65.
- van Marle G, Dobbe JC, Gultyaev AP, Luytjes W, Spaan WJ, Snijder EJ. Arterivirus discontinuous mRNA transcription is guided by base pairing between sense and antisense transcription-regulating sequences. Proc Natl Acad Sci U S A 1999 Oct 12;96(21):12056-61.
- van Marle G, van Dinten LC, Spaan WJ, Luytjes W, Snijder EJ. Characterization of an equine arteritis virus replicase mutant defective in subgenomic mRNA synthesis. J Virol 1999 Jul;73(7):5274-81.
- van Dinten LC, Rensen S, Gorbalenya AE, Snijder EJ. Proteolytic processing of the open reading frame 1b-encoded part of arterivirus replicase is mediated by nsp4 serine protease and Is essential for virus replication. J Virol 1999 Mar;73(3):2027-37.
- Maeda A, An S, Makino S. Importance of coronavirus negative-strand genomic RNA synthesis prior to subgenomic RNA transcription. Virus Res 1998 Sep;57(1):35-42.
- van der Meer Y, van Tol H, Locker JK, Snijder EJ. ORF1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex. J Virol 1998 Aug;72(8):6689-98.
- Lai MM, Cavanagh D. The molecular biology of coronaviruses. Adv Virus Res 1997;48:1-100.
- van Dinten LC, den Boon JA, Wassenaar AL, Spaan WJ, Snijder EJ. An infectious arterivirus cDNA clone: identification of a replicase point mutation that abolishes discontinuous mRNA transcription. Proc Natl Acad Sci U S A 1997 Feb 4;94(3):991-6.
- den Boon JA, Kleijnen MF, Spaan WJ, Snijder EJ. Equine arteritis virus subgenomic mRNA synthesis: analysis of leader-body junctions and replicative-form RNAs. J Virol 1996 Jul;70(7):4291-8.
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