Gene mapping of infectious bovine rhinotracheitis viral DNA genome

Simard, C; Nadon, Francine; Séguin, Cécile; Laboissiere, Sylvie; Trudel, Michel

doi:10.1007/bf01310703

Cited by 14 publications

(13 citation statements)

References 25 publications

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“…The UL25 ORF is located within fragment A at positions 60602r62398 (in the leftward orientation). The 4299 bp ApaI-BamHI fragment, representing the region 58917-63216 of the viral genome and shown in an expanded view, was subcloned from pKS/Ahd carrying the HindIII fragment A (Simard et al, 1990) to generate pKS/BHV58-63kApa-Bam. This recombinant plasmid was used as illustrated to create the recombinant eukaryotic expression vector pRetroTET-OFF/UL25 for the development of a cell line inducibly expressing UL25.…”

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Implication of the product of the bovine herpesvirus type 1 UL25 gene in capsid assembly

Desloges

Simard

2003

Journal of General Virology

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The UL25 ORF of bovine herpesvirus type 1 (BHV-1) was demonstrated recently to represent a gene encoding a 63 kDa viral protein. To investigate the role of this gene in virus replication, a BHV-1 UL25 deletion mutant was constructed. Although the UL25 mutant synthesizes late viral proteins and viral DNA, it fails to produce virus progeny in cells that do not express the UL25 gene, demonstrating that the UL25 protein is essential for the replicative cycle of BHV-1. Moreover, Southern blotting analyses of HindIII-digested DNA from infected non-complementing cells probed with the leftward terminal fragment of the BHV-1 linear genome revealed that the cleavage of the viral DNA produced is not impaired. However, the packaging of this cleaved DNA is compromised severely, since only few full C capsids were observed in infected non-complementing cells by transmission electron microscopy.Similar to herpes simplex virus type 1 (HSV-1), bovine herpesvirus type 1 (BHV-1) is a member of the subfamily Alphaherpesvirinae. Infection of host cells by these two viruses is similar, beginning with the entry of the nucleocapsid into the cytoplasm and translocation of the viral DNA into the nucleus. After expression of viral proteins, assembly and release of new virions occurs (Roizman & Sears, 1996).During the replication process, cleavage and packaging of viral DNA are tightly linked functions at least requiring the following seven genes in HSV-1: UL6, UL15, UL17, UL25, UL28, UL32 and UL33 (Poon & Roizman, 1993;Taus et al., 1998;McNab et al., 1998; Tengelsen et al., 1993;Addison et al., 1990;Lamberti & Weller, 1996al-Kobaisi et al., 1991). Viruses mutated in one of these genes synthesize near-wild-type levels of viral DNA but fail to cleave and package concatemeric DNA into capsids, resulting in an accumulation of B (intermediate) capsids in infected cells, with the exception of the UL25 mutant, which was shown to cleave newly synthesized viral DNA (McNab et al., 1998). Moreover, in contrast to the latter study where no packaging of matur DNA was observed, Stow (2001) reported that a significant amount of DNA synthesized by the UL25 mutant was packaged stably into capsids.To propagate a BHV-1 deletion mutant with a potentially lethal mutation in the UL25 gene to study the function of this gene, it was necessary to develop a complementing cell line expressing the UL25 protein in trans. For this purpose, RK13 cells (rabbit kidney cells, ATCC CCL-377) were stably transfected with 2 mg ScaI-linearized pRetroTET-OFF/UL25, encoding the complete UL25 ORF under the control of a tetracycline-regulated promoter (Fig. 1). A selected cell line grown in the absence of doxycycline, a tetracycline derivative compound (Gossen et al., 1995), expressed a 63 kDa protein that reacted specifically with the UL25 antiserum (results not shown; Desloges et al., 2001). This polypeptide represents the full-length product of UL25, since its size correlated with that of the protein observed in BHV-1-infected cells (Desloges et al., 2001). This cell line, named...

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Implication of the product of the bovine herpesvirus type 1 UL25 gene in capsid assembly

Desloges

Simard

2003

Journal of General Virology

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“…The latter is bounded by inverted repeat sequences (Rs) and can invert its orientation relative to UL, resulting in the existence of two isomeric forms (Farley et al, 1981). The genome encodes more than 40 polypeptides (Metzler et al, 1985;Misra et al, 1981), 20 of which have been located in discrete genomic regions using translation of mRNA hybrid-selected to individual HindlII DNA fragments (Simard et al, 1990). In particular, coding sequences of an abundant polypeptide (hereafter named VP8) with an apparent Mr of 94K have been assigned to the small 3.7 kbp HindlII fragment M. At present, gene coding…”

Section: Introductionmentioning

confidence: 99%

“…(subtype 1.1, Metzler et al, 1985) based on restriction endonuclease (Simard et al, 1991) and serological analyses. Virus was grown in confluent monolayers of an ovine kidney (OK) cell line as previously described (Trudel et al, 1987).…”

Section: Introductionmentioning

confidence: 99%

“…Poly(A) + mRNA was isolated as described (Simard et al, 1990) from ceils at 0, 6, 12, 18 and 24 h post-infection (p.i.) with BHV-1.…”

Section: Introductionmentioning

confidence: 99%

“…Virus was grown in confluent monolayers of an ovine kidney (OK) cell line as previously described (Trudel et al, 1987). Extracellular virions were concentrated by ultrafiltration (Trudel & Payment, 1980) and viral DNA was purified as reported (Simard et al, 1990).…”

Section: Introductionmentioning

confidence: 99%

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Characterization and transcript mapping of a bovine herpesvirus type 1 gene encoding a polypeptide homologous to the herpes simplex virus type 1 major tegument proteins VP13/14

Laboissiere

Trudel²,

Simard³

1992

Journal of General Virology

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Mapping of the bovine herpesvirus 1 glycoprotein C promoter region and its specific transactivation by the viral BICP27 gene product

Hamel

Simard

2003

Archives of Virology

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We investigated the cis-acting sequences of the promoter regulating the gene encoding the glycoprotein C (gC) of bovine herpesvirus 1 (BoHV-1), a gene of the gamma1 class. S1 nuclease protection assays revealed that gC transcription initiated predominantly at position C18281 of the viral genome, corresponding to 21 and 56 bases downstream from putative TATA-like and CAAT boxes, respectively. To map the gC promoter (pgC), we measured the ability of a series of nested deletions of the region +71 to -1155 with respect to the major gC transcriptional start site, to drive expression of the firefly luciferase (Luc) reporter gene following transient transfection of a cell host, in the absence as well as in the presence of viral factors expressed in trans. We show that the minimal pgC sequences required to drive maximal BoHV-1 independent expression was within the region +71 to -76 which harbours the putative TATA and CAAT boxes, the mRNA start site, and the complete 5' transcript leader sequence. This small pgC fragment was highly trans-activated by the co-expression of the BoHV-1-encoded BICP27 protein, but not BICP0 nor BTIF. In contrast, the pgC fragment spanning the region +71 to -1155 was only minimally trans-activated by BICP27, but substantially stimulated by BICP0. These findings thus suggest that gC gene regulation may involve the combined action of several viral transactivators.

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Gene mapping of infectious bovine rhinotracheitis viral DNA genome

Cited by 14 publications

References 25 publications

Implication of the product of the bovine herpesvirus type 1 UL25 gene in capsid assembly

Implication of the product of the bovine herpesvirus type 1 UL25 gene in capsid assembly

Characterization and transcript mapping of a bovine herpesvirus type 1 gene encoding a polypeptide homologous to the herpes simplex virus type 1 major tegument proteins VP13/14

Mapping of the bovine herpesvirus 1 glycoprotein C promoter region and its specific transactivation by the viral BICP27 gene product

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