DOI: 10.1128/jvi.74.6.2770-2776.2000. The Kinetics of VP5 mRNA Expression Is Not Critical for Viral Replication in Cultured Cells

The Kinetics of VP5 mRNA Expression Is Not Critical for Viral Replication in Cultured Cells

10.1128/jvi.74.6.2770-2776.2000

Crossref journal-article

American Society for Microbiology

Journal of Virology (235)

Abstract

ABSTRACTWe generated recombinant viruses in which the kinetics of expression of the leaky-late VP5 mRNA was altered. We then analyzed the effect of such alterations on viral replication in cultured cells. The VP5 promoter and leader sequences from positions −36 to +20, containing the TATA box and an initiator element, were deleted and replaced with a strong early (dUTPase), an equal-strength leaky-late (VP16), or a strict-late (UL38) promoter. We found that recombinant viruses containing the dUTPase promoter inserted in the VP5 locus expressed VP5-encoding mRNA with early kinetics, while virus with the UL38 promoter inserted expressed such mRNA with strict-late kinetics. Further, in spite of differences in its functional architecture, the VP16 promoter fully substituted for the VP5 promoter. Western blot analysis demonstrated that the amounts of VP5 capsid protein produced by the recombinant viruses differed somewhat; however, on complementing C32 and noncomplementing Vero cells, such viruses replicated to titers equivalent to those of the rescued wild-type virus controls. Multistep virus growth in mouse embryo fibroblasts, rabbit skin cells, and Vero cells also demonstrated equivalent replication efficiencies for both recombinant and wild-type viruses. Further, recombinant viruses did not show any impairment in their ability to replicate on serum-starved or quiescent human lung fibroblasts. We conclude that the kinetics of the essential VP5 mRNA expression is not critical for viral replication in cultured cells.

Bibliography

Lieu, P. T., & Wagner, E. K. (2000). The Kinetics of VP5 mRNA Expression Is Not Critical for Viral Replication in Cultured Cells. Journal of Virology, 74(6), 2770â2776.

Authors 2

Pauline T. Lieu (first)
Edward K. Wagner (additional)

References 38 Referenced 12

10.1128/jvi.63.5.2260-2269.1989
Blair E. D. Snowden B. W. Comparative analysis of the parameters that regulate expression from promoters of two late HSV-1 gene products Herpesvirus transcription and its regulation. Wagner E. K. 1991 181 206 CRC Press Boca Raton Fla
10.1128/jvi.67.3.1357-1364.1993
10.1128/jvi.65.2.769-786.1991
Flanagan W. M. Wagner E. K. A bi-functional reporter plasmid for the simultaneous transient expression assay of two herpes simplex virus promoters.Virus Genes 1 1987 61 71 (10.1007/BF00125686) / Virus Genes / A bi-functional reporter plasmid for the simultaneous transient expression assay of two herpes simplex virus promoters by Flanagan W. M. (1987)
10.1073/pnas.83.2.256
10.1128/jvi.68.12.7774-7789.1994
10.1128/jvi.67.9.5098-5108.1993
Homa F. L. Krikos A. Glorioso J. C. Levine M. Functional analysis of regulatory regions controlling strict late HSV gene expression Herpesvirus transcription and its regulation. Wagner E. K. 1991 207 222 CRC Press Boca Raton Fla
10.1128/jvi.67.9.5109-5116.1993
10.1128/jvi.68.9.5738-5747.1994
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10.1128/JVI.73.10.8843-8847.1999
10.1099/0022-1317-72-9-2311
10.1128/JVI.72.3.2463-2473.1998
10.1023/A:1008108121028
McGeoch D. J. Correlation between HSV-1 DNA sequence and viral transcription maps Herpesvirus transcription and its regulation. Wagner E. K. 1991 29 48 CRC Press Boca Raton Fla
10.1002/j.1460-2075.1988.tb03320.x
Pande N. T. Functional analysis of two early gene promoters of herpes simplex virus type 1. Ph.D. dissertation. 1997 University of California Irvine.
10.1006/viro.1997.9005
Roizman B. HSV gene functions: what have we learned that could be generally applicable to its near and distant cousins? Acta Virol. 43 1999 75 80 / Acta Virol. / HSV gene functions: what have we learned that could be generally applicable to its near and distant cousins? by Roizman B. (1999)
Roizman B. Tognon M. Restriction endonuclease patterns of herpes simplex virus DNA: application to diagnosis and molecular epidemiology.Curr. Top. Microbiol. Immunol. 104 1983 273 286 / Curr. Top. Microbiol. Immunol. / Restriction endonuclease patterns of herpes simplex virus DNA: application to diagnosis and molecular epidemiology by Roizman B. (1983)
10.1128/mcb.3.11.2028-2044.1983
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10.3109/00016489209137502
10.1128/jvi.69.6.3690-3703.1995
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Wagner E. K. Transcription patterns in herpes simplex virus infections Advances in viral oncology Klein G. III 1983 239 270 Raven Press New York, N.Y / Advances in viral oncology / Transcription patterns in herpes simplex virus infections by Wagner E. K. (1983)
Wagner E. K. Herpes simplex virus—molecular biology Encyclopedia of virology. Webster R. G. Granoff A. 1999 686 697 Academic Press London United Kingdom (10.1006/rwvi.1999.0123)
Wagner E. K. Anderson K. P. Costa R. H. Devi-Rao G. B. Gaylord B. Holland L. E. Stringer J. R. Tribble L. Isolation and characterization of HSV-1 mRNA Herpesvirus DNA: recent studies on the internal organization and replication of the viral genome. Becker Y. 1981 45 68 Martinus Nijhoff B.V. The Hague The Netherlands (10.1007/978-94-015-6897-5_4)
Wagner E. K. Costa R. H. Devi-Rao G. B. Draper K. Frink R. J. Hall L. Rice M. Steinhart W. Herpesvirus mRNA Developments in molecular virology Becker Y. VI 1985 79 100 Martinus Nijhoff The Hague, The Netherlands / Developments in molecular virology / Herpesvirus mRNA by Wagner E. K. (1985)
Wagner E. K. Guzowski J. F. Singh J. Transcription of the herpes simplex virus genome during productive and latent infection Progress in nucleic acid research and molecular biology. Cohen W. E. Moldave K. 1995 123 168 Academic Press San Diego Calif (10.1016/S0079-6603(08)60878-8)
10.1006/meth.1998.0648
10.1016/0022-2836(87)90487-6

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 1:55 a.m.)
Deposited	1 year, 8 months ago (Jan. 5, 2024, 1:48 p.m.)
Indexed	11 months, 3 weeks ago (Sept. 13, 2024, 12:41 p.m.)
Issued	25 years, 5 months ago (March 15, 2000)
Published	25 years, 5 months ago (March 15, 2000)
Published Print	25 years, 5 months ago (March 15, 2000)

Funders 0

None

BibTeX

@article{Lieu_2000, title={The Kinetics of VP5 mRNA Expression Is Not Critical for Viral Replication in Cultured Cells}, volume={74}, ISSN={1098-5514}, url={http://dx.doi.org/10.1128/jvi.74.6.2770-2776.2000}, DOI={10.1128/jvi.74.6.2770-2776.2000}, number={6}, journal={Journal of Virology}, publisher={American Society for Microbiology}, author={Lieu, Pauline T. and Wagner, Edward K.}, year={2000}, month=mar, pages={2770–2776} }

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