Pogliano K J, Beckwith J
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.
J Bacteriol. 1994 Feb;176(3):804-14. doi: 10.1128/jb.176.3.804-814.1994.
The secD operon of Escherichia coli is required for the efficient export of proteins. We have characterized this operon, and found that, in addition to secD and secF, it contains the upstream gene yajC, but not the genes queA or tgt, in contrast to previous reports. An analysis of yajC mutations constructed in vitro and recombined onto the chromosome indicates that yajC is neither essential nor a sec gene. The secD operon is not induced in response to either secretion defects or temperature changes. TnphoA fusions have been used to analyze the topology of SecD in the inner membrane; the protein contains six transmembrane stretches and a large periplasmic domain. TnphoA fusions to SecD and SecF have also been recombined onto the chromosome and used to determine the level of these proteins within the cell. Our results indicate that there are fewer than 30 SecD and SecF molecules per cell.
大肠杆菌的secD操纵子是蛋白质高效输出所必需的。我们已对该操纵子进行了表征,发现除了secD和secF外,它还包含上游基因yajC,但与之前的报道相反,它不包含queA或tgt基因。对体外构建并重组到染色体上的yajC突变进行分析表明,yajC既不是必需基因也不是sec基因。secD操纵子不会因分泌缺陷或温度变化而被诱导。TnphoA融合蛋白已被用于分析内膜中SecD的拓扑结构;该蛋白包含六个跨膜区段和一个大的周质结构域。与SecD和SecF的TnphoA融合蛋白也已重组到染色体上,并用于确定这些蛋白在细胞内的水平。我们的结果表明,每个细胞中SecD和SecF分子少于30个。