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首页> 外文期刊>Molecular and Cellular Biology >The conserved carboxy-terminal domain of Saccharomyces cerevisiae TFIID is sufficient to support normal cell growth.
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The conserved carboxy-terminal domain of Saccharomyces cerevisiae TFIID is sufficient to support normal cell growth.

机译：酿酒酵母TFIID的保守的羧基末端结构域足以支持正常细胞生长。

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We have examined the structure-function relationships of TFIID through in vivo complementation tests. A yeast strain was constructed which lacked the chromosomal copy of SPT15, the gene encoding TFIID, and was therefore dependent on a functional plasmid-borne wild-type copy of this gene for viability. By using the plasmid shuffle technique, the plasmid-borne wild-type TFIID gene was replaced with a family of plasmids containing a series of systematically mutated TFIID genes. These various forms of TFIID were expressed from three different promoter contexts of different strengths, and the ability of each mutant form of TFIID to complement our chromosomal TFIID null allele was assessed. We found that the first 61 amino acid residues of TFIID are totally dispensable for vegetative cell growth, since yeast strains containing this deleted form of TFIID grow at wild-type rates. Amino-terminally deleted TFIID was further shown to be able to function normally in vivo by virtue of its ability both to promote accurate transcription initiation from a large number of different genes and to interact efficiently with the Gal4 protein to activate transcription of GAL1 with essentially wild-type kinetics. Any deletion removing sequences from within the conserved carboxy-terminal region of S. cerevisiae TFIID was lethal. Further, the exact sequence of the conserved carboxy-terminal portion of the molecule is critical for function, since of several heterologous TFIID homologs tested, only the highly related Schizosaccharomyces pombe gene could complement our S. cerevisiae TFIID null mutant. Taken together, these data indicate that all important functional domains of TFIID appear to lie in its carboxy-terminal 179 amino acid residues. The significance of these findings regarding TFIID function are discussed.
机译：我们已经通过体内互补测试检查了TFIID的结构-功能关系。构建了一种酵母菌株，该菌株缺少SPT15（编码TFIID的基因）的染色体拷贝，因此依赖于该基因的功能性质粒携带的野生型拷贝来获得生存力。通过使用质粒改组技术，将质粒携带的野生型TFIID基因替换为包含一系列系统突变的TFIID基因的质粒家族。从不同强度的三个不同启动子背景表达了这些不同形式的TFIID，并评估了每种突变形式的TFIID补充我们的染色体TFIID无效等位基因的能力。我们发现TFIID的前61个氨基酸残基对于营养细胞的生长是完全可有可无的，因为含有这种缺失形式的TFIID的酵母菌株以野生型速率生长。进一步显示，氨基末端缺失的TFIID能够促进大量不同基因的准确转录起始，并能有效地与Gal4蛋白相互作用，从而激活GAL1的转录，从而具有体内正常功能。型动力学。来自酿酒酵母TFIID的保守的羧基末端区域内的任何删除去除序列都是致命的。此外，分子保守的羧基末端部分的确切序列对于功能至关重要，因为测试了几个异源TFIID同源物，只有高度相关的裂殖酵母pombe基因才能与我们的酿酒酵母TFIID空突变体互补。总而言之，这些数据表明TFIID的所有重要功能域似乎都位于其羧基末端的179个氨基酸残基中。讨论了有关TFIID功能的这些发现的意义。

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