Roeder G S, Rose A B, Pearlman R E
Proc Natl Acad Sci U S A. 1985 Aug;82(16):5428-32. doi: 10.1073/pnas.82.16.5428.
The his4-917 mutation of yeast results from the insertion of a Ty element, Ty917, into the 5' regulatory region of the HIS4 gene. Ty917 prevents HIS4 transcription, thus rendering the cell histidine requiring. Recombination between Ty917 and a Ty element elsewhere in the yeast genome can result in the replacement of part or all of the Ty917 element by sequences from the Ty element. Recombinant derivatives display a variety of phenotypes including His-, weakly His+, and strongly His+. In most of the His+ derivatives, the expression of HIS4 is controlled by genes at the mating type locus. To identify the Ty sequences important in controlling the expression of an adjacent gene, we used Ty elements that have different effects on gene expression to construct hybrid Ty elements in vitro. The effects of these hybrid elements on HIS4 expression were examined. These experiments indicate that the critical sequence differences between Ty elements that permit HIS4 expression and those that prevent its expression lie in the rightmost (HIS4-proximal) 730 base pairs of the element. The DNA sequence of this region was determined for three elements: Ty917, which prevents HIS4 expression; Ty917(467), which confers a weak His+ phenotype; and Ty917(480), which confers a strong His+ phenotype. Within this region, Ty917(467) differs from Ty917 by a single base-pair change that is in the internal (epsilon) region of the Ty element. Ty917(480) differs from Ty917 by this same base-pair change and by 10 changes in the terminal delta sequence. The sequence change common to Ty917(467) and Ty917(480) lies in a region of the Ty element that is homologous to the simian virus 40 enhancer of transcription.
酵母的his4 - 917突变是由于一个Ty元件Ty917插入到HIS4基因的5'调控区所致。Ty917阻止HIS4转录,从而使细胞需要组氨酸。Ty917与酵母基因组中其他位置的Ty元件之间的重组可导致Ty917元件的部分或全部被来自该Ty元件的序列所取代。重组衍生物表现出多种表型,包括组氨酸缺陷型(His -)、弱组氨酸阳性型(weakly His +)和强组氨酸阳性型(strongly His +)。在大多数组氨酸阳性衍生物中,HIS4的表达受交配型位点基因的控制。为了鉴定在控制相邻基因表达中起重要作用的Ty序列,我们使用了对基因表达有不同影响的Ty元件在体外构建杂交Ty元件。检测了这些杂交元件对HIS4表达的影响。这些实验表明,允许HIS4表达的Ty元件与阻止其表达的Ty元件之间的关键序列差异位于该元件最右侧(靠近HIS4)的730个碱基对中。测定了三个元件该区域的DNA序列:阻止HIS4表达的Ty917;赋予弱组氨酸阳性表型的Ty917(467);以及赋予强组氨酸阳性表型的Ty917(480)。在该区域内,Ty917(467)与Ty917的差异在于Ty元件内部(ε)区域的一个单碱基对变化。Ty917(480)与Ty917的差异在于这个相同的碱基对变化以及末端δ序列中的10个变化。Ty917(467)和Ty917(480)共有的序列变化位于Ty元件中与猴病毒40转录增强子同源的区域。