Efficient activation of transcription in yeast by the BPV1 E2 protein.
The full-length gene product encoded by the E2 open reading frame (ORF) of bovine papillomavirus type 1 (BPV1) is a transcriptional transactivator. It is believed to mediate its effect on the BPV1 long control region (LCR) by binding to motifs with the consensus sequence ACCN6GGT. The minimal functional cis active site, called the E2 response element (E2RE), in mammalian cells comprises two copies of this motif. Here we have shown that E2 can function in Saccharomyces cerevisiae by placing an E2RE upstream of a synthetic yeast assay promoter which consists of a TATA motif and an mRNA initiation site, spaced correctly. This E2RE-minimal promoter is only transcriptionally active in the presence of E2 protein and the resulting mRNA is initiated at the authentic start site. This is the first report of a mammalian viral transactivator functioning in yeast. The level of activation by E2 via the E2RE was the same as observed with the highly efficient authentic PGK promoter where the upstream activation sequence is composed of three distinct elements. Furthermore a single E2 motif which is insufficient in mammalian cells as an activation site was as efficiently utilized in yeast as the E2RE (2 motifs). Previous studies have shown that mammalian cellular activators can function in yeast and our data now extend this to viral-specific activators. Our data indicate however that while the mechanism of transactivation is broadly conserved there may be significant differences at the detailed level.
Stanway
C A
CA
Department of Biochemistry, Oxford University, UK.
Sowden
M P
MP
Wilson
L E
LE
Kingsman
A J
AJ
Kingsman
S M
SM
D001483:Base Sequence; D052688:Bovine papillomavirus 1; D004271:DNA, Fungal / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000494:pharmacology*; D004742:Enhancer Elements, Genetic; D007370:Interferon Type I / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects*; D014764:Viral Proteins / Q000378:metabolism / Q000494:pharmacology*
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C051550:E2 protein, Bovine papillomavirus; D007370:Interferon Type I; D012333:RNA, Messenger; D014764:Viral Proteins