The herpesvirus saimiri open reading frame (ORF) 50 (Rta) protein contains an at hook required for binding to the ORF 50 response element in delayed-early promoters. Academic Article uri icon

Overview

MeSH

  • Amino Acid Sequence
  • Animals
  • Cell Line
  • DNA-Binding Proteins
  • Gene Deletion
  • Gene Expression Regulation, Viral
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed

MeSH Major

  • AT-Hook Motifs
  • Bacterial Proteins
  • Heat-Shock Proteins
  • Herpesvirus 2, Saimiriine
  • Promoter Regions, Genetic
  • Response Elements
  • Transcriptional Activation

abstract

  • The herpesvirus saimiri open reading frame (ORF) 50 encodes two proteins, which activate transcription directly, following interactions with delayed-early (DE) promoters containing a specific motif. In this report, we demonstrate that ORF 50 contains a DNA binding domain that has homology to an AT hook DNA binding motif. Deletion analysis of this domain reduces ORF 50-mediated transactivation of the DE ORF 6 and ORF 57 promoters by 100 and 90%, respectively. Furthermore, gel retardation experiments demonstrated that the AT hook motif is required for binding the ORF 50 response element in the promoters of DE genes. Single site-directed mutagenesis of the AT hook revealed that mutation of the glycine residue at position 408 to an alanine reduces ORF 50 transactivation of the ORF 57 promoter by 40%. Moreover, the mutation of multiple basic residues in conjunction with the glycine residue within the core element of the AT hook abolishes ORF 50-mediated transactivation. In addition, p50GFPDeltaAT-hook is capable of functioning as a trans-dominant mutant, leading to a reduction in virus production of approximately 50% compared to that for wild-type ORF 50.

publication date

  • May 2004

has subject area

  • AT-Hook Motifs
  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins
  • Cell Line
  • DNA-Binding Proteins
  • Gene Deletion
  • Gene Expression Regulation, Viral
  • Heat-Shock Proteins
  • Herpesvirus 2, Saimiriine
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Promoter Regions, Genetic
  • Response Elements
  • Transcriptional Activation

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC387665

PubMed ID

  • 15078979

Additional Document Info

start page

  • 4936

end page

  • 4942

volume

  • 78

number

  • 9