Chambers A, Stanway C, Kingsman A J, Kingsman S M
Department of Biochemistry, University of Oxford, UK.
Nucleic Acids Res. 1988 Sep 12;16(17):8245-60. doi: 10.1093/nar/16.17.8245.
The UAS (upstream activator sequence) of the yeast PGK gene contains a transcriptional activator domain located between bases -479 and -402 upstream from the initiating ATG. This region of the UAS contains three direct repeats of the sequence 5'CTTCC3'. The roles in transcriptional activation of these repeats and other sequences within the activator domain were investigated. When short regions containing the repeats were removed PGK expression was considerably reduced, the magnitude of the effect depended upon which CTTCC block was absent. Sequences between -473 and -458 which did not contain a CTTCC block were also shown to be necessary for high levels of PGK expression. A DNA fragment containing activator sequences up to -473 was shown to interact specifically in vitro with a yeast nuclear protein extract. DNase I footprinting identified a protected region between -473 and -458 and single base changes in DNase I sensitivity at the CTTCC repeats.
酵母磷酸甘油酸激酶（PGK）基因的上游激活序列（UAS）含有一个转录激活结构域，位于起始ATG上游-479至-402碱基之间。UAS的这一区域包含序列5'CTTCC3'的三个直接重复序列。对这些重复序列以及激活结构域内其他序列在转录激活中的作用进行了研究。当去除包含重复序列的短区域时，PGK表达显著降低，其影响程度取决于缺失的是哪个CTTCC片段。-473至-458之间不包含CTTCC片段的序列对于高水平的PGK表达也是必需的。一个包含至-473的激活序列的DNA片段在体外与酵母核蛋白提取物发生特异性相互作用。DNA酶I足迹法确定了-473至-458之间的一个受保护区域以及CTTCC重复序列处DNA酶I敏感性的单碱基变化。
