Transposable element-mediated enhancement of gene expression in Saccharomyces cerevisiae involves sequence-specific binding of a trans-acting factor.
In our studies on the regulation of adjacent-gene expression by Ty sequences, we demonstrated that a single-base-pair change (T-A----C-G) in the epsilon sequence of Ty917-derived elements is primarily responsible for enhancement of beta-galactosidase expression from lacZ fusion plasmids. Using an electrophoretic gel mobility assay, we showed that the same base pair transition is required for binding of a trans-acting factor, TyBF, from crude cell extracts in vitro. We identified the site of TyBF binding and determined the guanine nucleotide contact sites required for TyBF interaction. We propose that TyBF binding to cis-acting Ty2 sequences activates adjacent-gene transcription.
Goel
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Department of Biology, York University, Toronto, Ontario, Canada.
Pearlman
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D001483:Base Sequence; D001667:Binding, Competitive; D003227:Conjugation, Genetic; D004247:DNA / Q000032:analysis / Q000378:metabolism; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D005656:Fungal Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation; D008745:Methylation; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D014157:Transcription Factors; D004247:DNA