Shimotsu H, Henner D J
J Bacteriol. 1986 Oct;168(1):380-8. doi: 10.1128/jb.168.1.380-388.1986.
In Bacillus subtilis, the extracellular enzyme levansurcrase is synthesized in the presence of sucrose. A termination structure between the transcription start site and the structural gene was the apparent site for regulation by sucrose of transcription into the structural gene. Sequence analysis of the sacB leader region from two strains constitutive for levansucrase synthesis showed a single base change in the stem of this termination structure. This single base change also led to the constitutive synthesis of a sacB'-'lacZ fusion, whereas the wild-type sacB'-'lacZ fusion was induced by the addition of sucrose. S1 nuclease mapping of sacB transcripts with probes labeled either within the termination structure or 3' to the termination structure showed that sucrose did not increase the number of transcripts extending into the termination structure; however, sucrose did increase the number of transcripts extending past the termination structure. Two pleiotropic mutations which affect the expression of levansucrase, sacQ36 hyperproducing [sacQ36(Hy)] and sacU32(Hy), were separately introduced into the strain carrying the sacB'-'lacZ fusion. These mutations each increased the expression levels of the sacB'-'lacZ fusion. S1 mapping showed increased levels of transcript initiating at the sacB promoter in strains with the sacQ36(Hy) and sacU32(Hy) mutations. This increased transcription appeared to be independent of the sucrose-regulated transcription termination, suggesting the existence of at least two different mechanisms for the regulation of sacB expression.
在枯草芽孢杆菌中,胞外酶果聚糖蔗糖酶在蔗糖存在的情况下合成。转录起始位点与结构基因之间的一个终止结构是蔗糖对转录进入结构基因进行调控的明显位点。对两株果聚糖蔗糖酶合成组成型菌株的sacB前导区进行序列分析,结果显示该终止结构的茎部存在一个单碱基变化。这一单碱基变化还导致了sacB'-'lacZ融合基因的组成型合成,而野生型sacB'-'lacZ融合基因是通过添加蔗糖诱导表达的。用在终止结构内或终止结构3'端标记的探针进行sacB转录本的S1核酸酶图谱分析表明,蔗糖并没有增加延伸到终止结构内的转录本数量;然而,蔗糖确实增加了延伸超过终止结构的转录本数量。分别将影响果聚糖蔗糖酶表达的两个多效性突变,即超产的sacQ36 [sacQ36(Hy)] 和sacU32(Hy),导入携带sacB'-'lacZ融合基因的菌株中。这些突变各自提高了sacB'-'lacZ融合基因的表达水平。S1图谱分析表明,在具有sacQ36(Hy)和sacU32(Hy)突变的菌株中,从sacB启动子起始的转录本水平有所增加。这种转录增加似乎与蔗糖调节的转录终止无关,这表明至少存在两种不同的sacB表达调控机制。