Homologous recognition of a promoter domain common to the MSV LTR and the HSV tk gene

Barbara J. Graves, Peter F. Johnson, Steven L. McKnight

Research output: Contribution to journalArticlepeer-review

367 Scopus citations

Abstract

We have partially purified, from rat liver, a nuclear protein fraction with sequence-specific affinity to a promoter domain shared by the herpesvirus tk gene and the Moloney murine sarcoma virus LTR. For both promoters, the protein-binding domain occurs roughly 80 bp upstream of the mRNA cap site and harbors the pentanucleotide sequence 5′-CCAAT-3′. The MSV LTR pentanucleotide occurs on the coding strand in an orientation pointing toward the retroviral transcription unit. The HSV tk pentanucleotide occurs on the non-coding strand in an orientation pointing away from the gene. Assays in microinjected frog oocytes and transfected mouse L cells indicate that equivalent point mutations, introduced at each residue of the CAT pentanucleotide of each promoter, lead to similar changes in promoter activity. Furthermore, they similarly alter binding of the nuclear protein fraction. Surprisingly, a C to G transversion at the first residue of the CAT pentanucleotide, which severely impairs the activity of both promoters, appears to increase affinity of the CAT binding protein.

Original languageEnglish (US)
Pages (from-to)565-576
Number of pages12
JournalCell
Volume44
Issue number4
DOIs
StatePublished - Feb 28 1986

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

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