Puyet A, Ibáñez A M, Espinosa M
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
J Biol Chem. 1993 Dec 5;268(34):25402-8.
The gene encoding a transcriptional repressor of the maltosaccharide utilization operons of the Gram-positive bacterium Streptococcus pneumoniae (malR) has been cloned and sequenced. The genetic structure of the locus reveals the presence of an upstream gene necessary for growth on maltotetraose medium (malA). The phenotype of malR- and malA- mutants obtained by interruption of the coding regions suggests that both genes could belong to the same transcription unit. Two copies of a DNA motif consisting of three conserved regions of 59, 42, and 49 nucleotides were found located upstream and downstream of the malA-malR putative operon. These DNA structures are almost identical to the reported Box sequences associated with several genes of S. pneumoniae. The protein encoded by malR was visualized and partially purified after selective expression in Escherichia coli, whereas the product of malA was identified in vitro. The amino acid sequence of MalR displays similarities with the Lac and Gal family of repressors. The highest similarities were found when comparing MalR with the E. coli MalI repressor, which is related with an indirect induction pathway of the maltose regulon. The significance of these similarities is discussed in terms of the possible evolutionary pathways followed by structural and regulatory genes of sugar utilization systems in bacteria.
编码革兰氏阳性细菌肺炎链球菌麦芽糖利用操纵子转录阻遏物的基因(malR)已被克隆和测序。该基因座的遗传结构表明,在麦芽四糖培养基上生长所必需的一个上游基因(malA)存在。通过中断编码区获得的malR和malA突变体的表型表明,这两个基因可能属于同一个转录单元。在假定的malA-malR操纵子的上游和下游发现了由59、42和49个核苷酸的三个保守区域组成的DNA基序的两个拷贝。这些DNA结构与报道的与肺炎链球菌几个基因相关的Box序列几乎相同。在大肠杆菌中选择性表达后,对malR编码的蛋白质进行了可视化和部分纯化,而malA的产物在体外得到了鉴定。MalR的氨基酸序列与Lac和Gal阻遏物家族具有相似性。在将MalR与大肠杆菌MalI阻遏物进行比较时发现了最高的相似性,MalI阻遏物与麦芽糖调节子的间接诱导途径有关。从细菌中糖利用系统的结构和调节基因可能遵循的进化途径方面讨论了这些相似性的意义。