Characterization of Berne virus genomic and messenger RNAs.
Abstract: From 380S particles of Berne virus (proposed family Toroviridae) one species of polyadenylated RNA was isolated. Using agarose gel electrophoresis its length was estimated as 20 kb or greater. When assayed under hypertonic transfection conditions genomic RNA was found to be infectious; RNase treatment destroyed the infectivity. The positive polarity of the molecule was confirmed by filter spot hybridization using cDNA prepared against poly(A)-selected RNA from infected cells. In embryonic mule skin cells infected with Berne virus the presence of five virus-specific, polyadenylated RNA species of 7.5, 2.1, 1.4, 0.8 and at least 20 kb was demonstrated. In vitro translation of the 7.5, 2.1 and 0.8 kb RNAs followed by immunoprecipitation showed that they encode a 151K product (possibly the precursor to the peplomer proteins), the envelope protein and the nucleocapsid protein, respectively.
Publication Date: 1988-09-01 PubMed ID: 3411297DOI: 10.1099/0022-1317-69-9-2135Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article deals with understanding the characteristics of various RNA types associated with Berne virus, a proposed member of the Toroviridae family, and their subsequent roles in the virus’s life cycle.
Isolation and Characterization of Berne Virus RNA
- The study began with the isolation of a particular type of polyadenylated RNA from the 380S particles of Berne virus. Polyadenylated RNA refers to RNA molecules that have a string of adenylic acid residues at the end, which is characteristic of most eukaryotic messenger RNAs (mRNAs).
- This RNA was estimated to have a length of 20 kilobases (kb) or more, determined via agarose gel electrophoresis, a technique commonly used for the separation and analysis of macromolecules such as DNA and RNA.
- The researchers observed that under hypertonic transfection conditions, the genomic RNA was found to be infectious, i.e., capable of causing disease. RNase treatment, which involves the application of a type of enzyme that degrades RNA, eliminated this infectivity, further validating its role.
Confirming the Polarity and Identifying Other Virus-Specific RNAs
- The positive polarity of the isolated RNA molecule was confirmed using filter spot hybridization with complementary DNA (cDNA), prepared against poly(A)-selected RNA from infected cells. In the context of RNA, positive polarity denotes that the RNA is of the same sense as mRNA and can be directly translated into proteins.
- Existing alongside the main 20 kb RNA, the researchers found five other virus-specific polyadenylated RNA species of varying lengths (7.5 kb, 2.1 kb, 1.4 kb, 0.8 kb, and at least 20 kb) in embryonic mule skin cells infected with Berne virus.
Determining Protein Encodings of the RNAs
- Through in vitro translation and subsequent immunoprecipitation, a process used to isolate a specific protein out of a mixture, it was determined that the 7.5 kb, 2.1 kb, and 0.8 kb RNAs encode for a 151K product (possibly a precursor to the peplomer proteins), the envelope protein, and the nucleocapsid protein, respectively. These proteins play crucial roles in the structure and activities of the virus.
- The results of this study provide descriptive information about the genomic and messenger RNAs of the Berne virus, which can be instrumental in understanding the life cycle and pathogenic capabilities of the virus.
Cite This Article
APA
Snijder EJ, Ederveen J, Spaan WJ, Weiss M, Horzinek MC.
(1988).
Characterization of Berne virus genomic and messenger RNAs.
J Gen Virol, 69 ( Pt 9), 2135-2144.
https://doi.org/10.1099/0022-1317-69-9-2135 Publication
Researcher Affiliations
- Institute of Virology, Veterinary Faculty, State University of Utrecht, The Netherlands.
MeSH Terms
- Animals
- Horses / microbiology
- Molecular Weight
- Protein Biosynthesis
- RNA Viruses / genetics
- RNA, Messenger / genetics
- RNA, Viral / genetics
- Viral Proteins / genetics
- Virus Replication
Citations
This article has been cited 15 times.- Shi Z, Wang W, Chen C, Zhang X, Wang J, Xu Z, Lan Y. First report and genetic characterization of bovine torovirus in diarrhoeic calves in China.. BMC Vet Res 2020 Aug 5;16(1):272.
- de Vries AAF, Horzinek MC, Rottier PJM, de Groot RJ. The Genome Organization of the Nidovirales: Similarities and Differences between Arteri-, Toro-, and Coronaviruses.. Semin Virol 1997 Feb;8(1):33-47.
- Petric M. VI, 1. Epidemiology of toroviruses.. Perspect Med Virol 2003;9:625-632.
- Dervas E, Hepojoki J, Laimbacher A, Romero-Palomo F, Jelinek C, Keller S, Smura T, Hepojoki S, Kipar A, Hetzel U. Nidovirus-Associated Proliferative Pneumonia in the Green Tree Python (Morelia viridis).. J Virol 2017 Nov 1;91(21).
- Koopmans M, Monroe SS, Coffield LM, Zaki SR. Optimization of extraction and PCR amplification of RNA extracts from paraffin-embedded tissue in different fixatives.. J Virol Methods 1993 Jul;43(2):189-204.
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- Snijder EJ, den Boon JA, Bredenbeek PJ, Horzinek MC, Rijnbrand R, Spaan WJ. The carboxyl-terminal part of the putative Berne virus polymerase is expressed by ribosomal frameshifting and contains sequence motifs which indicate that toro- and coronaviruses are evolutionarily related.. Nucleic Acids Res 1990 Aug 11;18(15):4535-42.
- Snijder EJ, Horzinek MC, Spaan WJ. A 3'-coterminal nested set of independently transcribed mRNAs is generated during Berne virus replication.. J Virol 1990 Jan;64(1):331-8.
- Snijder EJ, Den Boon JA, Spaan WJ, Weiss M, Horzinek MC. Primary structure and post-translational processing of the Berne virus peplomer protein.. Virology 1990 Oct;178(2):355-63.
- Koopmans M, Snijder EJ, Horzinek MC. cDNA probes for the diagnosis of bovine torovirus (Breda virus) infection.. J Clin Microbiol 1991 Mar;29(3):493-7.
- Den Boon JA, Snijder EJ, Locker JK, Horzinek MC, Rottier PJ. Another triple-spanning envelope protein among intracellularly budding RNA viruses: the torovirus E protein.. Virology 1991 Jun;182(2):655-63.
- Snijder EJ, den Boon JA, Horzinek MC, Spaan WJ. Comparison of the genome organization of toro- and coronaviruses: evidence for two nonhomologous RNA recombination events during Berne virus evolution.. Virology 1991 Jan;180(1):448-52.
- den Boon JA, Snijder EJ, Chirnside ED, de Vries AA, Horzinek MC, Spaan WJ. Equine arteritis virus is not a togavirus but belongs to the coronaviruslike superfamily.. J Virol 1991 Jun;65(6):2910-20.
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