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Related Experiment Videos

Long-term promoter activity during herpes simplex virus latency

J R Lokensgard1, D C Bloom, A T Dobson

  • 1Department of Microbiology and Immunology, UCLA School of Medicine 90024.

Journal of Virology
|November 1, 1994
PubMed
Summary

Herpes simplex virus type 1 (HSV-1) gene therapy vectors require promoters active during latency. The Moloney murine leukemia virus long terminal repeat (LTR) enabled sustained gene expression during latent infection, unlike other promoters tested.

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Area of Science:

  • Molecular Virology
  • Gene Therapy
  • Neuroscience

Background:

  • Directing foreign gene expression from herpes simplex virus type 1 (HSV-1) is crucial for gene therapy applications.
  • Understanding neuronal transcription factors and their variation across neuronal populations is limited.
  • The mechanism behind the continuous activity of the latency-associated transcript (LAT) promoter during HSV-1 latency remains unclear.

Purpose of the Study:

  • To investigate promoter activity from HSV-1 during acute and latent infections in neurons.
  • To identify elements responsible for sustained gene expression during HSV-1 latency.

Main Methods:

  • Constructed seven recombinant HSV-1 viruses with different promoter-lacZ reporter gene constructs at the glycoprotein C locus.
  • Evaluated beta-galactosidase expression during acute and latent infections in murine dorsal root ganglia.

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  • Compared the activity of murine metallothionein (MT1), phosphoglycerate kinase, Moloney murine leukemia virus long terminal repeat (LTR), and HSV LAT core promoter constructs.

    • Main Results:

    • Promoter activities varied widely during acute infection; MT1, phosphoglycerate kinase, Moloney murine leukemia virus LTR, and LAT were active acutely but not latently.
    • Incorporating LAT upstream transcription factor binding sites into MT1 and LTR promoters (LAT-MT1, LAT-LTR) enhanced acute-phase expression.
    • Only the LAT-LTR construct maintained transcriptional activity after the establishment of latency, indicating the Moloney murine leukemia virus LTR confers latency-associated activity.

    Conclusions:

    • The Moloney murine leukemia virus LTR contains a DNA element that prevents promoter inactivation during HSV-1 latency.
    • The upstream LAT promoter region does not contain the HSV long-term-expression element necessary for sustained activity during latency.
    • HSV long-term-expression function is likely provided by a region distinct from that mediating high-level acute-phase neuronal expression.