tailor-made zinc-finger transcription factors activate flo11 gene expression with phenotypic consequences in the yeast saccharomyces cerevisiae特制的锌指转录因子激活flo11基因表达的表型影响酵母酿酒酵母.pdfVIP

Tailor-Made Zinc-Finger Transcription Factors Activate FLO11 Gene Expression with Phenotypic Consequences in the Yeast Saccharomyces cerevisiae 1,2 1 3 2 2¤ 2 Jia-Ching Shieh *, Yu-Che Cheng , Mao-Chang Su , Michael Moore , Yen Choo , Aaron Klug 1 Department of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan, 2 Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom, 3 Department of Otorhinolaryngology–Head and Neck Surgery, Chung Shan Medical University Hospital, Taichung, Taiwan Cys His zinc fingers are eukaryotic DNA-binding motifs, capable of distinguishing different DNA sequences, and are suitable 2 2 for engineering artificial transcription factors. In this work, we used the budding yeast Saccharomyces cerevisiae to study the ability of tailor-made zinc finger proteins to activate the expression of the FLO11 gene, with phenotypic consequences. Two three-finger peptides were identified, recognizing sites from the 59 UTR of the FLO11 gene with nanomolar DNA-binding affinity. The three-finger domains and their combined six-finger motif, recognizing an 18-bp site, were fused to the activation domain of VP16 or VP64. These transcription factor constructs retained their DNA-binding ability, with the six-finger ones being the highest in affinity. However, when expressed in haploid yeast cells, only one three-finger recombinant transcription factor was able to activate the expression of FLO11 efficiently. Unlike in the wild-type, cells with such transcriptional activation displayed invasive growth and biofilm formation, without any requirement for glucose depletion. The VP16 and VP64 domains appeared to act equally well in the activation of FLO11 expression, with comparable effects in phenotypic alteration. We co