Transcriptionally active immediate-early protein of pseudorabies virus binds to specific sites on class II gene promoters.
In the presence of partially purified pseudorabies virus immediate-early protein, multiple sites of DNase I protection were observed on the adenovirus major late and human hsp 70 promoters. Southwestern (DNA-protein blot) analysis demonstrated that the immediate-early protein bound directly to the sequences contained in these sites. These sequences share only limited homology, differ in their affinities for the immediate-early protein, and are located at different positions on these two promoters. In addition, the site-specific binding of a temperature-sensitive immediate-early protein was eliminated by the same heat treatment which eliminates its transcriptional activating function, whereas the binding of the wild-type protein was unaffected by heat treatment. Thus, site-specific binding requires a functionally active immediate-early protein. Furthermore, immediate-early-protein-dependent in vitro transcription from the major late promoter was preferentially inhibited by oligonucleotides which are homologous to the high-affinity binding sites on the major late or hsp 70 promoters. These observations suggest that transcriptional stimulation by the immediate-early protein involves binding to cis-acting elements.
Cromlish
W A
WA
Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021-6399.
Abmayr
S M
SM
Workman
J L
JL
Horikoshi
M
M
Roeder
R G
RG
D001483:Base Sequence; D001667:Binding, Competitive; D015139:Blotting, Southern; D015153:Blotting, Western; D004268:DNA-Binding Proteins / Q000502:physiology*; D003850:Deoxyribonuclease I / Q000494:pharmacology; D005786:Gene Expression Regulation; D006360:Heat-Shock Proteins / Q000235:genetics; D011558:Herpesvirus 1, Suid / Q000235:genetics*; D017874:Immediate-Early Proteins; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014157:Transcription Factors / Q000502:physiology*; D014764:Viral Proteins / Q000502:physiology*
D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; D017874:Immediate-Early Proteins; D015345:Oligonucleotide Probes; D014157:Transcription Factors; D014764:Viral Proteins; C047583:pseudorabies virus immediate early proteins; D003850:Deoxyribonuclease I