IDENTIFICATION OF SEQUENCES RESPONSIBLE FOR TRANSCRIPTIONAL REGULATION OF THE STRONGLY EXPRESSED METHANOL OXIDASE-ENCODING GENE IN HANSENULA-POLYMORPHA
IDENTIFICATION OF SEQUENCES RESPONSIBLE FOR TRANSCRIPTIONAL REGULATION OF THE STRONGLY EXPRESSED METHANOL OXIDASE-ENCODING GENE IN HANSENULA-POLYMORPHA
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DOI：
10.1016/0378-1119(94)90520-7
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发表时间：
1994-02-11
期刊：
影响因子：
3.5
通讯作者：
HOLLENBERG, CP
中科院分区：
文献类型：
--
作者：
GODECKE, S;ECKART, M;HOLLENBERG, CP
甲基营养酵母菌由于其高度调节甲醇代谢和过氧化物体的生物发生而成为深入研究的对象。我们对酵母MOX基因的5‘调控区进行了研究，该基因编码甲醇代谢的关键酶--过氧化体甲醇氧化酶。这个调控严密的约1.5kb的酵母启动子非常大，在诱导条件下也具有显著的强度，属于迄今描述的最强的酵母启动子。缺失分析显示了一个复杂的启动子结构，该结构由几个对报告基因表达具有正负调控作用的序列元件组成，并且元件之间存在明显的协同作用。通过凝胶滞留实验和DNase I足迹实验检测了几种因子在体外的特异性结合。根据缺失数据，可以确定两个结合位点为上游激活序列(UAS1和UAS2)，一个结合位点为上游抑制序列(URS1)。
The methylotrophic yeasts have been the subject of intensive studies, because of their highly regulated methanol metabolism and the biogenesis of peroxisomes. We investigated the 5' regulatory region of the MOX gene from the yeast, Hansemula polymorpha, encoding the peroxisomal methanol oxidase, the key enzyme of methanol metabolism. This tightly regulated yeast promoter of approximately 1.5 kb is unusually large, and also of remarkable strength under inducing conditions, belonging to the strongest yeast promoters yet described. Deletion analyses revealed a complex promoter structure composed of several sequence elements with positive and negative regulatory effects on reporter gene expression and a pronounced cooperation between the elements. Specific binding of several factors was detected in vitro by gel retardation and DNase I footprinting experiments. On the basis of deletion data, two binding sites could be identified as upstream activation sequences (UAS1 and UAS2) and one binding site as an upstream repressing sequence (URS1).