Crutz A M, Steinmetz M
Laboratoire de Génétique des Microorganismes, Centre National de la Recherche Scientifique (URA 537), Thiverval-Grignon, France.
J Bacteriol. 1992 Oct;174(19):6087-95. doi: 10.1128/jb.174.19.6087-6095.1992.
The adjacent sacX and sacY genes are involved in sucrose induction of the Bacillus subtilis sacB gene by an antitermination mechanism. sacB, encoding the exoenzyme levansucrase, is also subject to regulation by the DegS-DegU signalling system. Using sacXY'-lacZ and sacX'-lacZ fusions, we show that the transcription of the sacX and sacY genes is both inducible by sucrose and regulated by DegU. sacX and sacY appear to constitute an operon, since the deletion of the sacX leader region abolished the expression of a sacXY'-lacZ fusion. The degU-dependent promoter was located by deletion analysis and reverse transcriptase mapping 300 nucleotides upstream from the sacX initiator codon. Sucrose induction of the sacX'-lacZ fusion requires either SacY or the homologous SacT antiterminator, which is involved in sucrose induction of the intracellular sucrase gene (sacPA operon). Sequence analysis of the sacX leader region revealed (20 nucleotides downstream from the transcription start site) a putative binding site for these regulators; however, no structure resembling a rho-independent terminator could be found overlapping this site, unlike the situation for sacPA and sacB. Deletion of a segment of the leader region located 100 nucleotides downstream from this site led to constitutive expression of the sacXY'-lacZ and sacX'-lacZ fusions. These results suggest that the mechanism of sucrose induction of sacXY is different from that of sacPA and sacB.
相邻的sacX和sacY基因通过抗终止机制参与枯草芽孢杆菌sacB基因的蔗糖诱导。编码外切酶果聚糖蔗糖酶的sacB也受DegS - DegU信号系统调控。利用sacXY'-lacZ和sacX'-lacZ融合体,我们发现sacX和sacY基因的转录既受蔗糖诱导,也受DegU调控。sacX和sacY似乎构成一个操纵子,因为sacX前导区的缺失消除了sacXY'-lacZ融合体的表达。通过缺失分析和逆转录酶定位,将依赖degU的启动子定位在sacX起始密码子上游300个核苷酸处。sacX'-lacZ融合体的蔗糖诱导需要SacY或同源的SacT抗终止子,SacT参与细胞内蔗糖酶基因(sacPA操纵子)的蔗糖诱导。sacX前导区的序列分析揭示(转录起始位点下游20个核苷酸处)这些调节因子的一个假定结合位点;然而,与sacPA和sacB的情况不同,在该位点重叠处未发现类似不依赖于rho的终止子的结构。删除该位点下游100个核苷酸处的一段前导区导致sacXY'-lacZ和sacX'-lacZ融合体的组成型表达。这些结果表明sacXY的蔗糖诱导机制与sacPA和sacB不同。