Identification of herpes simplex virus DNA sequences which encode a trans-acting polypeptide responsible for stimulation of immediate early transcription

We investigated the feasibility of using retroviruses as vectors for transferring DNA sequences into animal cells. The thymidine kinase (tk) gene of herpes simplex virus was chosen as a convenient model. The internal BamHI fragments of a DNA clone of Moloney leukemia virus (MLV) were replaced with a purified BamHI DNA segment containing the tk gene. Chimeric genomes were created carrying the tk insert in both orientations relative to the MLV sequence. Each was transfected into TK- cells along with MLV helper virus, and TK+ colonies were obtained by selection in the presence of hypoxanthine, aminopterin, and thymidine (HAT). Virus collected from TK+-transformed, MLV producer cells passed the TK+ phenotype to TK- cells. Nonproducer cells were isolated, and TK+ transducing virus was subsequently rescued from them. The chimeric virus showed single-hit kinetics in infections. Virion and cellular RNA and cellular DNA from infected cells were all shown to contain sequences which hybridized to both MLV- and tk-specific probes. The sizes of these sequences were consistent with those predicted for the chimeric virus. In all respects studied, the chimeric MLV-tk virus behaved like known replication-defective retroviruses. These experiments suggest great general applicability of retroviruses as eucaryotic vectors.

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Trans-activation of Herpes Simplex virus type 1 immediate early genes is specifically inhibited by human recombinant interferons

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(89)90011-9 ◽

1989 ◽

Vol 159 (2) ◽

pp. 439-444 ◽

Cited By ~ 3

Author(s):

P.Raniero De Stasio ◽

Milton W. Taylor

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Immediate Early Genes ◽

Immediate Early ◽

Early Genes ◽

Simplex Virus

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Herpes Simplex Virus ICP27 Activation of Stress Kinases JNK and p38

Journal of Virology ◽

10.1128/jvi.79.13.8348-8360.2005 ◽

2005 ◽

Vol 79 (13) ◽

pp. 8348-8360 ◽

Cited By ~ 63

Author(s):

Danna Hargett ◽

Tim McLean ◽

Steven L. Bachenheimer

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Viral Gene Expression ◽

Immediate Early ◽

Wild Type Virus ◽

Viral Gene ◽

Functional Domain ◽

Viral Protein Synthesis ◽

Immediate Early Proteins ◽

Simplex Virus

ABSTRACT We previously reported that herpes simplex virus type 1 (HSV-1) can activate the stress-activated protein kinases (SAPKs) p38 and JNK. In the present study, we undertook a comprehensive and comparative analysis of the requirements for viral protein synthesis in the activation of JNK and p38. Infection with the UL36 mutant tsB7 or with UV-irradiated virus indicated that both JNK and p38 activation required viral gene expression. Cycloheximide reversal or phosphonoacetic acid treatment of wild-type virus-infected cells as well as infection with the ICP4 mutant vi13 indicated that only the immediate-early class of viral proteins were required for SAPK activation. Infection with ICP4, ICP27, or ICP0 mutant viruses indicated that only ICP27 was necessary. Additionally, we determined that in the context of virus infection ICP27 was sufficient for SAPK activation and activation of the p38 targets Mnk1 and MK2 by infecting with mutants deleted for various combinations of immediate-early proteins. Specifically, the d100 (0−/4−) and d103 (4−/22−/47−) mutants activated p38 and JNK, while the d106 (4−/22−/27−/47−) and d107 (4−/27−) mutants did not. Finally, infections with a series of ICP27 mutants demonstrated that the functional domain of ICP27 required for activation was located in the region encompassing amino acids 20 to 65 near the N terminus of the protein and that the C-terminal transactivation activity of ICP27 was not necessary.

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Herpes Simplex Virus ICP27 Is Required for Virus-Induced Stabilization of the ARE-Containing IEX-1 mRNA Encoded by the Human IER3 Gene

Journal of Virology ◽

10.1128/jvi.01216-06 ◽

2006 ◽

Vol 80 (19) ◽

pp. 9720-9729 ◽

Cited By ~ 28

Author(s):

Jennifer A. Corcoran ◽

Wei-Li Hsu ◽

James R. Smiley

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Mapk Pathway ◽

Immediate Early ◽

Productive Infection ◽

Mrna Stabilization ◽

Early Protein ◽

Host Shutoff ◽

Simplex Virus ◽

Hsv 1

ABSTRACT Herpes simplex virus (HSV) stifles cellular gene expression during productive infection of permissive cells, thereby diminishing host responses to infection. Host shutoff is achieved largely through the complementary actions of two viral proteins, ICP27 and virion host shutoff (vhs), that inhibit cellular mRNA biogenesis and trigger global mRNA decay, respectively. Although most cellular mRNAs are thus depleted, some instead increase in abundance after infection; perhaps surprisingly, some of these contain AU-rich instability elements (AREs) in their 3′-untranslated regions. ARE-containing mRNAs normally undergo rapid decay; however, their stability can increase in response to signals such as cytokines and virus infection that activate the p38/MK2 mitogen-activated protein kinase (MAPK) pathway. We and others have shown that HSV infection stabilizes the ARE mRNA encoding the stress-inducible IEX-1 mRNA, and a previous report from another laboratory has suggested vhs is responsible for this effect. However, we now report that ICP27 is essential for IEX-1 mRNA stabilization whereas vhs plays little if any role. A recent report has documented that ICP27 activates the p38 MAPK pathway, and we detected a strong correlation between this activity and stabilization of IEX-1 mRNA by using a panel of HSV type 1 (HSV-1) isolates bearing an array of previously characterized ICP27 mutations. Furthermore, IEX-1 mRNA stabilization was abrogated by the p38 inhibitor SB203580. Taken together, these data indicate that the HSV-1 immediate-early protein ICP27 alters turnover of the ARE-containing message IEX-1 by activating p38. As many ARE mRNAs encode proinflammatory cytokines or other immediate-early response proteins, some of which may limit viral replication, it will be of great interest to determine if ICP27 mediates stabilization of many or all ARE-containing mRNAs.

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