Jarai G, Buxton F
Department of Biotechnology, Ciba-Geigy AG, Basel, Switzerland.
Curr Genet. 1994 Sep;26(3):238-44. doi: 10.1007/BF00309554.
Aspergillus niger secretes a number of enzymes, including proteases, into its culture fluid. The regulation of the two major acidic extracellular proteases, pepA and pepB, was investigated using Northern analyses. Our data suggest that the regulation of pepA and pepB expression occurs predominantly at the level of mRNA content and that, while they are regulated in a similar manner, differences are also clear in their expression. Both genes were found to be under complex regulatory control. The expression of the two genes could be turned off by the presence of good nitrogen or carbon sources in the media, and external protein sources did not induce expression of either gene under conditions of carbon and nitrogen repression. The pH of the medium also played a major role in their regulation as the expression of both genes was completely turned off under alkaline conditions, even when grown in media lacking good nitrogen and carbon sources but containing proteins. We isolated clones containing 5' non-coding sequences of the pepA gene from a lambda genomic library with a pepA specific probe. Analysis and comparison of the promoter sequences of the pepA and pepB genes revealed that both contain several putative AREA- and CREA-binding sites and they also share an 18-bp-long sequence which is 83% identical in these two genes.
黑曲霉会向其培养液中分泌多种酶,包括蛋白酶。使用Northern分析方法研究了两种主要的酸性细胞外蛋白酶pepA和pepB的调控机制。我们的数据表明,pepA和pepB表达的调控主要发生在mRNA含量水平,并且尽管它们以相似的方式受到调控,但在表达上也存在明显差异。发现这两个基因都受到复杂的调控。当培养基中存在优质氮源或碳源时,这两个基因的表达会被关闭,并且在碳氮抑制条件下,外部蛋白质源不会诱导任何一个基因的表达。培养基的pH值在它们的调控中也起着重要作用,因为即使在缺乏优质氮源和碳源但含有蛋白质的培养基中生长,在碱性条件下这两个基因的表达也会完全关闭。我们用pepA特异性探针从λ基因组文库中分离出含有pepA基因5'非编码序列的克隆。对pepA和pepB基因启动子序列的分析和比较表明,两者都含有几个假定的AREA和CREA结合位点,并且它们还共享一个18个碱基对长的序列,这两个基因中该序列的同源性为83%。