Mapping of 10 epitopes on bovine herpesvirus type 1 glycoproteins gl and glll

FitzPatrick, David R.; Redmondz, Mark J.; Attah-Poku, Samuel K.; Hurk, Sylvia van Drunen Littel‐van den; Babiuk, Lorne A.; Zamb, Timothy J.

doi:10.1016/0042-6822(90)90239-n

Cited by 34 publications

(36 citation statements)

References 23 publications

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“…Two expression vectors, pRSVgI and pRSDneogI, containing the gI-coding sequence under the control of the Rous sarcoma virus long terminal repeat enhancerpromoter were constructed as previously described (Fitzpatrick et al, 1988(Fitzpatrick et al, , 1990. The vector pRSDneogI contains all of the elements of pRSVgI but with the addition of the neomycin resistance gene, under the control of the simian virus (SV40) enhancer-early promoter.…”

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confidence: 99%

“…, 1984), and processed with an avidin biotin-enhanced immunoperoxidase staining procedure (Fitzpatrick et al, 1988). (Fitzpatrick, 1989). Babiuk, 1985).…”

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confidence: 99%

“…Constitutive expression of r e c o m b i n a n t gI in transfected M D B K cells was obtained using a strategy similar to that employed previously for L M T K -cells (Fitzpatrick et al, 1988), but using modified Rous sarcoma virus-based expression vectors which are more efficient than SV40-based vectors in transfected bovine cells (Fitzpatrick, 1989). Cloned cell lines derived from transfections with the vector p R S D n e o g I consistently exhibited higher levels of g!…”

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confidence: 99%

“…This was not detectable by immunocytochemistry (data not shown) but was demonstrated using flow cytometric analysis (Fig. 2) of transfected cells labelled with a gI-specific MAb followed by a fluorescein isothiocyanate-conjugated goat anti-mouse immunoglobulin (Fitzpatrick, 1989).…”

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confidence: 99%

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Expression of Bovine Herpesvirus Type 1 Glycoprotein gI in Transfected Bovine Cells Induces Spontaneous Cell Fusion

FitzPatrick

Zamb²,

Babiuk

1990

Journal of General Virology

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Bovine MDBK cells were transfected with Rous sarcoma virus-based vectors for constitutive expression of the bovine herpesvirus type 1 (BHV-1) glycoprotein, gI. Cell lines stably expressing recombinant gI were cloned and characterized. Recombinant gI was localized intracellularly, predominantly in a perinuclear region, and on the cell surface. Cells expressing gI exhibited spontaneous polykaryon formation, thus confirming the fusogenic activity described previously in gI-expressing transfected murine LMTK-cells. The recombinant form of gI synthesized in transfected MDBK cells was similar in Mr to the form expressed in BHV-l-infected MDBK cells, unlike the recombinant form ofgI expressed by LMTK-cells which is deficient in N-linked glycosylation. It was concluded that cell fusion associated with the expression of BHV-1 gI in transfected mammalian cells is a reproducible phenomenon in a number of cell types and is not due to species-specific factors or expression of abnormally glycosylated gI. Cell fusion is a useful in vitro marker for gI function and may contribute to the spread of BHV-1 infections in vivo.

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confidence: 99%

“…, 1984), and processed with an avidin biotin-enhanced immunoperoxidase staining procedure (Fitzpatrick et al, 1988). (Fitzpatrick, 1989). Babiuk, 1985).…”

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confidence: 99%

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confidence: 99%

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confidence: 99%

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Expression of Bovine Herpesvirus Type 1 Glycoprotein gI in Transfected Bovine Cells Induces Spontaneous Cell Fusion

FitzPatrick

Zamb²,

Babiuk

1990

Journal of General Virology

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“…Previous studies have suggested that BHV-1 a t t a c h m e n t to permissive cells is m e d i a t e d by the envelope glycoproteins gI, gIII and gIV (Liang et al, 1991), whereas viral penetration of the host cells is m e d i a t e d by gI and gIV (Hughes et al, 1988;Fitzpatrick et al, 1990). It has been established that initial a t t a c h m e n t o f the virus to permissive cells is via the interaction of gIII (and gI and gIV in the absence o f gIII) with heparin-like moieties on the cell surface (Okazaki et al, 1991).…”

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Identification of a putative cellular receptor for bovine herpesvirus 1

Thaker

Stine²,

Zamb³

et al. 1994

Journal of General Virology

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Functional Analysis of the Transmembrane Anchor Region of Bovine Herpesvirus 1 Glycoprotein gB

Hurk

Liang

et al. 1997

Virology

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In herpesviruses, homologues of glycoprotein B (gB) are essential membrane proteins which are involved in fusion. However, there is no clear evidence regarding the location of the fusogenic domain on gB. By using bovine herpesvirus 1 (BHV-1) as a model, we studied the relationship between the structure and the fusogenic activity of gB. This was achieved by expressing genes of different gB derivatives containing specific truncations at the end of segments 2 or 3 of the transmembrane region in Madin-Darby bovine kidney cells under the control of the bovine heat-shock protein hsp70A gene promoter. All expressed gB products were structurally similar to authentic gB. One truncated form of gB, gBt, which contains residues 1-763, was efficiently secreted. However, gBtM (residues 1-807), which includes the first two segments at the carboxyl terminus, showed unstable retention on the cell surface, whereas gBtMA (residues 1 829), which contains all three membrane-spanning segments, was mostly intracellularly retained with some unstable surface anchorage. Another truncated gB, gBtDAF, which has gB residues 1-763 (gBt) and a human decay-accelerating factor (DAF) carboxyl tail, was also expressed. The DAF fragment provided a signal for the addition of a glycosyl phosphatidylinositol-based membrane anchor, which could target the gBt chimeric protein on the cell membrane. Immunofluorescence staining and pulse-chase kinetic studies support the theory that gBtM, gBtMA, and gBtDAF are retained on nuclear and cellular membranes via different segments of the transmembrane region or the DAF fragment, respectively. For the cells expressing gBt or gBtM, no cell fusion was observed, whereas cells expressing gBtMA clearly showed fusion. However, in gBtDAF cells, the overexpression and cellular accumulation of recombinant gB products did not cause fusion either, which supports our contention that the fusion phenomenon in gBtMA cells is caused by the fusogenic activity of the expressed gBtMA. With the help of sequence analysis, our results indicate that segment 2 of the transmembrane anchor region might be a fusogenic domain, whereas the real anchor is segment 3.

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Mapping of 10 epitopes on bovine herpesvirus type 1 glycoproteins gl and glll

Cited by 34 publications

References 23 publications

Expression of Bovine Herpesvirus Type 1 Glycoprotein gI in Transfected Bovine Cells Induces Spontaneous Cell Fusion

Expression of Bovine Herpesvirus Type 1 Glycoprotein gI in Transfected Bovine Cells Induces Spontaneous Cell Fusion

Identification of a putative cellular receptor for bovine herpesvirus 1

Functional Analysis of the Transmembrane Anchor Region of Bovine Herpesvirus 1 Glycoprotein gB

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