Analysis of a Herpes Simplex Virus 2 Fragment from the Open Reading Frame of the Large Subunit of Ribonucleotide Reductase with Transcriptional Regulatory Activity

C. Jones, F. Zhu, K. R. Dhanwada

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Expression of herpes simplex virus 2 (HSV-2)-encoded ribonucleotide reductase (RR) is required for growth in nondividing cells. The functional enzyme is composed of a large and a small subunit. In virus-infected cells, RR is expressed temporally as a delayed early protein. However, the promoter regulatory region of the large subunit can function as an immediate early promoter in transient transfection assays, suggesting that expression may be quite complex. In this study, a 95-bp fragment derived from the open reading frame of the large subunit of RR (RR-A) functioned as a silencer when placed adjacent to a heterologous promoter. If the fragment was placed distal to the promoter, repression was relieved and in human keratinocytes promoter activity was consistently higher than control constructs. Exonuclease III protection assays revealed that nuclear factors from human keratinocytes as well as other primate cells specifically bind to this fragment. A 30-bp motif containing a consensus SP-1 binding site and an alternating Pu/Py element was protected in all cell lines. These results suggest that a 95-bp fragment in the open reading frame of HSV-2 RR-A plays a role in regulating viral gene expression.

Original languageEnglish (US)
Pages (from-to)127-137
Number of pages11
JournalDNA and Cell Biology
Volume12
Issue number2
DOIs
StatePublished - Mar 1993

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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