Analysis of the in vitro translation products of the equine herpesvirus type 1 immediate early mRNA.
Abstract: Equine herpesvirus type 1 (EHV-1) gene expression is coordinately regulated in an alpha, beta, gamma fashion. Viral alpha gene products include a 6.0-kb immediate early (IE) mRNA species (W. L. Gray et al., 1987, Virology 158, 79-87) and at least four closely related IE polypeptides (IEPs) (G.B. Caughman et al., 1985, Virology 145, 49-61). In this report, we describe results obtained from a series of in vitro translation experiments which were performed in an effort to characterize the IEPs and identify the mechanism by which individual IE protein species are generated. Our data indicate that a family of IEPs is generated in vitro from the 6.0-kb mRNA size class and that these IEPs correspond in overall size and antigenicity to those synthesized in infected cells. Using time-course/pulse-chase analyses, we show that production of three of the major IEPs [IE1' (193 kDa), IE3' (166 kDa), and IE4' (130 kDA)] occurs concomitantly, that none of these protein species can be chased completely into another, and that at least two additional minor species appear to be processed following synthesis. Finally, we show that the 6.0-kb mRNA species isolated during early or late stages of the infection cycle can be translated to yield all of the major IE proteins, indicating that production of the family of IEPs is not dependent upon accumulation of the IE mRNA which occurs during a cycloheximide blocked infection cycle. The implications of these findings are discussed as they relate to the origin and production of the IEPs both in vivo and in vitro.
Publication Date: 1988-10-01 PubMed ID: 2845657DOI: 10.1016/0042-6822(88)90516-8Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research paper discusses a study on the gene expression of Equine herpesvirus type 1 (EHV-1) and the role of the virus’s immediate early (IE) mRNA and its resulting proteins (IEPs), conducted through a series of in vitro translation experiments.
Overview and Aim
- The study aimed at understanding the role of IE mRNA and how its translation leads to the generation of different protein species known as IEPs in EHV-1.
- The researchers intended to investigate the nature of these IEPs and the mechanism through which they are generated.
Procedure and Findings
- In vitro translation experiments were conducted to analyse the translation products of the IE mRNA.
- The results showed that a family of IEPs are indeed generated from IE mRNA, and that these proteins corresponded with the ones synthesized within infected cells.
- Through time-course and pulse-chase analyses, it was highlighted that three primary IEPs were produced simultaneously and that these individual proteins couldn’t be fully converted into one another.
- The study also found evidence of at least two additional minor protein species that seemed to emerge following the synthesis process.
Key Conclusions
- The 6.0-kb mRNA could be translated at both early and late phases of the viral infection cycle into all major IEPs, negating the dependence on IE mRNA accumulation during an infection cycle blocked by cycloheximide (an antifungal medication).
- The findings provide crucial insights on the origin and production of IEPs both in vivo (inside the body) and in vitro (outside the body in a controlled environment).
Implications
- By understanding the generation and nature of these viral proteins, it could eventually open possibilities for developing new therapeutic strategies and treatments against EHV-1.
- Such findings can also contribute to enhancing our general understanding of virus biology and virus-host interaction mechanisms.
Cite This Article
APA
Robertson AT, Caughman GB, Gray WL, Baumann RP, Staczek J, O'Callaghan DJ.
(1988).
Analysis of the in vitro translation products of the equine herpesvirus type 1 immediate early mRNA.
Virology, 166(2), 451-462.
https://doi.org/10.1016/0042-6822(88)90516-8 Publication
Researcher Affiliations
- Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130-3932.
MeSH Terms
- Blotting, Northern
- Gene Expression Regulation
- Genes, Overlapping
- Genes, Viral
- Herpesviridae / genetics
- Herpesvirus 1, Equid / genetics
- Molecular Weight
- Precipitin Tests
- Protein Biosynthesis
- Protein Processing, Post-Translational
- RNA, Messenger / genetics
- RNA, Viral / genetics
- Time Factors
- Viral Proteins / genetics
Grant Funding
- AI21996 / NIAID NIH HHS
- AI22001 / NIAID NIH HHS
- AI22894 / NIAID NIH HHS
Citations
This article has been cited 14 times.- Jang HK, Albrecht RA, Buczynski KA, Kim SK, Derbigny WA, O'Callaghan DJ. Mapping the sequences that mediate interaction of the equine herpesvirus 1 immediate-early protein and human TFIIB.. J Virol 2001 Nov;75(21):10219-30.
- Smith RH, Zhao Y, O'Callaghan DJ. The equine herpesvirus 1 (EHV-1) UL3 gene, an ICP27 homolog, is necessary for full activation of gene expression directed by an EHV-1 late promoter.. J Virol 1993 Feb;67(2):1105-9.
- Holden VR, Caughman GB, Zhao Y, Harty RN, O'Callaghan DJ. Identification and characterization of the ICP22 protein of equine herpesvirus 1.. J Virol 1994 Jul;68(7):4329-40.
- O'Callaghan DJ, Colle CF 3rd, Flowers CC, Smith RH, Benoit JN, Bigger CA. Identification and initial characterization of the IR6 protein of equine herpesvirus 1.. J Virol 1994 Sep;68(9):5351-64.
- Caughman GB, Lewis JB, Smith RH, Harty RN, O'Callaghan DJ. Detection and intracellular localization of equine herpesvirus 1 IR1 and IR2 gene products by using monoclonal antibodies.. J Virol 1995 May;69(5):3024-32.
- Zhao Y, Holden VR, Smith RH, O'Callaghan DJ. Regulatory function of the equine herpesvirus 1 ICP27 gene product.. J Virol 1995 May;69(5):2786-93.
- Harty RN, Colle CF, Grundy FJ, O'Callaghan DJ. Mapping the termini and intron of the spliced immediate-early transcript of equine herpesvirus 1.. J Virol 1989 Dec;63(12):5101-10.
- Wirth UV, Gunkel K, Engels M, Schwyzer M. Spatial and temporal distribution of bovine herpesvirus 1 transcripts.. J Virol 1989 Nov;63(11):4882-9.
- Baumann RP, Yalamanchili VR, O'Callaghan DJ. Functional mapping and DNA sequence of an equine herpesvirus 1 origin of replication.. J Virol 1989 Mar;63(3):1275-83.
- Hayes MK, Rock DL. Identification of a novel bovine herpesvirus type 1 immediate-early infected cell protein.. Arch Virol 1990;112(3-4):291-300.
- Wirth UV, Vogt B, Schwyzer M. The three major immediate-early transcripts of bovine herpesvirus 1 arise from two divergent and spliced transcription units.. J Virol 1991 Jan;65(1):195-205.
- Harty RN, O'Callaghan DJ. An early gene maps within and is 3' coterminal with the immediate-early gene of equine herpesvirus 1.. J Virol 1991 Jul;65(7):3829-38.
- Holden VR, Yalamanchili RR, Harty RN, O'Callaghan DJ. ICP22 homolog of equine herpesvirus 1: expression from early and late promoters.. J Virol 1992 Feb;66(2):664-73.
- Smith RH, Caughman GB, O'Callaghan DJ. Characterization of the regulatory functions of the equine herpesvirus 1 immediate-early gene product.. J Virol 1992 Feb;66(2):936-45.
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