Alexander D M, St John A C
Department of Biological Sciences, Rutgers, State University of New Jersey, Piscataway 08855-1059.
Mol Microbiol. 1994 Mar;11(6):1059-71. doi: 10.1111/j.1365-2958.1994.tb00383.x.
Escherichia coli induces the expression of more than 50 proteins in response to starvation for a carbon source. Strains MC7 (csi7::phoA) and MC19 (csi19::phoA) contain fusions of a signal peptide-deficient phoA reporter sequence to a csi (carbon starvation-inducible) gene. PhoA expression increased when these strains were deprived of a carbon source or entered stationary phase but did not when the cells were deprived of a nitrogen source or subjected to osmotic, oxidative or thermal stress. Mapping and sequence analysis of the cloned phoA fusions in strains MC7 and MC19 indicated that they had occurred in different locations within the same previously unidentified gene. The wild-type allele of this gene was cloned and the encoded protein was found to be a new lipoprotein. Therefore we propose to call this locus slp (starvation lipoprotein). The 22 kDa Slp protein is associated with the outer membrane fraction. The slp gene was located at 78.6 centisomes on the E. coli genetic map. The -10 and -35 regions upstream of the mRNA start site were characteristic of a sigma 70 promoter. The major transcript from this promoter was sufficiently large to contain slp sequences but not the downstream open reading frame. Induction of beta-galactosidase activity from a slp::lacZ translational fusion during carbon starvation or stationary phase was independent of cAMP, RpoS (KatF) and DnaK, all of which are known to affect the expression of certain starvation-inducible or stationary phase-inducible proteins.
大肠杆菌在碳源饥饿时会诱导50多种蛋白质的表达。菌株MC7(csi7::phoA)和MC19(csi19::phoA)含有信号肽缺陷型phoA报告序列与csi(碳饥饿诱导型)基因的融合体。当这些菌株被剥夺碳源或进入稳定期时,PhoA表达增加,但当细胞被剥夺氮源或受到渗透、氧化或热应激时则不会增加。对菌株MC7和MC19中克隆的phoA融合体进行定位和序列分析表明,它们发生在同一先前未鉴定基因的不同位置。克隆了该基因的野生型等位基因,发现编码的蛋白质是一种新的脂蛋白。因此,我们提议将这个基因座称为slp(饥饿脂蛋白)。22 kDa的Slp蛋白与外膜部分相关。slp基因位于大肠杆菌遗传图谱的78.6厘摩处。mRNA起始位点上游的-10和-35区域具有σ70启动子的特征。来自该启动子的主要转录本足够大,包含slp序列,但不包含下游的开放阅读框。在碳饥饿或稳定期,从slp::lacZ翻译融合体诱导β-半乳糖苷酶活性与cAMP、RpoS(KatF)和DnaK无关,已知这些都影响某些饥饿诱导型或稳定期诱导型蛋白质的表达。