Equine Herpesviruses: Biochemical studies on genomic structure, DI particles, oncogenic transformation and persistent infection

O’Callaghan, Dennis J.; Henry, Berch E.; Wharton, J. H.; Dauenhauer, Steven A.; Vance, Ralph B.; Staczek, John; Atherton, Sally S.; Robinson, Robin A.

doi:10.1007/978-94-015-6897-5_20

Cited by 28 publications

(29 citation statements)

References 35 publications

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“…Our studies on the nature of the EHV-1 defective interfering particles (DIP) capable of mediating a state of persistent infection (41,50) revealed that the DIP genome harbors repeated copies of a unique hybrid ORF that encodes the aminoterminal 196 aa of EICP22P and the carboxy-terminal 68 aa of EHV-1 EICP27P (13). This hybrid ORF, which encodes portions of two major EHV-1 early regulatory proteins, is abundantly expressed at both the mRNA and protein levels in cells infected with virus preparations containing DIP (13).…”

Section: Discussionmentioning

confidence: 99%

The EICP22 Protein of Equine Herpesvirus 1 Physically Interacts with the Immediate-Early Protein and with Itself To Form Dimers and Higher-Order Complexes

Derbigny

Kim

Caughman

et al. 2000

J Virol

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The EICP22 protein (EICP22P) of Equine herpesvirus 1 (EHV-1) is an early protein that functions synergistically with other EHV-1 regulatory proteins to transactivate the expression of early and late viral genes. We have previously identified EICP22P as an accessory regulatory protein that has the ability to enhance the transactivating properties and the sequence-specific DNA-binding activity of the EHV-1 immediate-early protein (IEP). In the present study, we identify EICP22P as a self-associating protein able to form dimers and higher-order complexes during infection. Studies with the yeast two-hybrid system also indicate that physical interactions occur between EICP22P and IEP and that EICP22P self-aggregates. Results from in vitro and in vivo coimmunoprecipitation experiments and glutathione S-transferase (GST) pull-down studies confirmed a direct protein-protein interaction between EICP22P and IEP as well as self-interactions of EICP22P.

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Section: Discussionmentioning

confidence: 99%

The EICP22 Protein of Equine Herpesvirus 1 Physically Interacts with the Immediate-Early Protein and with Itself To Form Dimers and Higher-Order Complexes

Derbigny

Kim

Caughman

et al. 2000

J Virol

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“…Procedures have been described for isolation of viral DNA (2,8,9), generation of DI particles (4,5) and monitoring for DI particles by density analysis (4), and restriction enzyme analysis ofviral DNA (1). EHV-1 DI-particle DNA was obtained from the supramolar fragment of Bgl II digests of DI-particle preparations (1). Plasmid pBR322 DNA (10) was extracted from large preparations of bacteria by the technique of Guerry et aL (11) and was banded by centrifugation in CsCl/ethidium bromide gradients (12).…”

Section: Methodsmentioning

confidence: 99%

“…Methods for restriction enzyme digestions, separation of DNA fragments by agarose gel electrophoresis, and isolation of individual DNA fragments have been described (1,2). EHV-1 and pBR322 (13) DNAs were digested separately to completion with BamHI or EcoRI.…”

Section: Methodsmentioning

confidence: 99%

“…Only the S region contains unique sequences flanked by inverted repetitive sequences (1) which allow the genome to exist in two isomeric forms (2). Upon repeated, high-multiplicity passage of EHV-1 in animals (3) or cell culture (4,5), defective interfering (DI) particles, whose genome is comprised of tandem repeat units originating from the S region (1,4), emerge. One striking consequence ofthe interference activity ofthese DI particles is that infection of permissive hamster embryo cells with viable preparations of these defective particles results in oncogenic transformation and establishment of persistent infection (1,6).…”

mentioning

confidence: 99%

“…Upon repeated, high-multiplicity passage of EHV-1 in animals (3) or cell culture (4,5), defective interfering (DI) particles, whose genome is comprised of tandem repeat units originating from the S region (1,4), emerge. One striking consequence ofthe interference activity ofthese DI particles is that infection of permissive hamster embryo cells with viable preparations of these defective particles results in oncogenic transformation and establishment of persistent infection (1,6). Because DNA sequences located in the L region appear to be associated with EHV-1 oncogensis (7), we have proposed a model in which these herpesvirus DI particles mediate oncogenic transformation indirectly by tempering cytocidal viral functions and thereby allowing oncogenic DNA sequences to be dominantly expressed (6).…”

mentioning

confidence: 99%

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Molecular cloning of equine herpesvirus type 1 DNA: analysis of standard and defective viral genomes and viral sequences in oncogenically transformed cells.

Robinson¹,

Tucker²,

Dauenhauer³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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Genomic DNA sequences of equine herpesvirus type 1 (EHV-1) have been cloned as BamHI and EcoRI restriction fragments into the plasmid pBR322 and propagated in Escherichia coli. With the exception of two EcoRI restriction fragments that reside in the S region of the viral genome, all of the cloned fragments demonstrated the same electrophoretic mobilities, restriction cleavage sites, and blot-hybridization patterns as did the parent fragments produced by BamHI or EcoRI digestion of virion DNA. The EcoRI J fragment and the BamHI E fragment of the L-region terminus were cloned after the addition of appropriate linker oligonucleotides. Fragments originating from each of the two isomeric forms of EHV-1 DNA were contained in this library of clones. Supramolar DNA fragments present only in the DNA of defective interfering (DI) particles of EHV-1 were generated from Bgl II digestion of DNA preparations enriched for EHV-1 DI particles and were cloned as Bgl II and EcoRI fragments into the plasmid vector. The cloned viral sequences represented in this defective genome mapped to the S region of EHV-1 DNA. Blothybridization analysis ofEHV-1 transformed and tumor cell DNAs with the cloned BamHI B fragment confirmed that subgenomic viral sequences are present and indicated that those sequences map to the viral genome between 0.32 and 0.43 map unit.The EHV41 genome is a 92-megadalton (MDal) DNA molecule comprised of L and S covalently linked regions. Only the S region contains unique sequences flanked by inverted repetitive sequences (1) which allow the genome to exist in two isomeric forms (2). Upon repeated, high-multiplicity passage of EHV-1 in animals (3) or cell culture (4, 5), defective interfering (DI) particles, whose genome is comprised of tandem repeat units originating from the S region (1, 4), emerge. One striking consequence ofthe interference activity ofthese DI particles is that infection of permissive hamster embryo cells with viable preparations of these defective particles results in oncogenic transformation and establishment of persistent infection (1, 6). Because DNA sequences located in the L region appear to be associated with EHV-1 oncogensis (7), we have proposed a model in which these herpesvirus DI particles mediate oncogenic transformation indirectly by tempering cytocidal viral functions and thereby allowing oncogenic DNA sequences to be dominantly expressed (6).One approach for providing ample quantities of viral DNA sequences necessary for biochemical analyses of virus-transformed cells is the insertion of viral DNA restriction enzyme fragments by recombinant DNA methods into plasmids for subsequent propagation in bacteria. In this report we describe the isolation and characteristics of cloned plasmids containing restriction fragments of EHVr1 standard and defective genomes and demonstrate the utilization of cloned viral DNA fragments to identify EHV-1 DNA sequences in oncogenically transformed and tumor cells. MATERIALS AND METHODSCells, Virus, and DNA. Procedures have been described for...

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The Equine Herpesviruses

1983

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Equine Herpesviruses: Biochemical studies on genomic structure, DI particles, oncogenic transformation and persistent infection

Cited by 28 publications

References 35 publications

The EICP22 Protein of Equine Herpesvirus 1 Physically Interacts with the Immediate-Early Protein and with Itself To Form Dimers and Higher-Order Complexes

The EICP22 Protein of Equine Herpesvirus 1 Physically Interacts with the Immediate-Early Protein and with Itself To Form Dimers and Higher-Order Complexes

Molecular cloning of equine herpesvirus type 1 DNA: analysis of standard and defective viral genomes and viral sequences in oncogenically transformed cells.

The Equine Herpesviruses

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