Frey Alexander D, Andersson Charlotte I J, Schmid Vinzenz H, Bülow Leif, Kallio Pauli T
Institute of Microbiology, ETH-Zürich, CH-8093 Zürich, Switzerland.
J Biotechnol. 2007 May 1;129(3):461-71. doi: 10.1016/j.jbiotec.2007.01.012. Epub 2007 Jan 25.
We have analyzed gene expression of Escherichia coli MG1655 expressing native and engineered bacterial globin proteins, in order to identify the molecular mechanisms leading to the improved phenotypical traits relative to control cells under oxygen-limited conditions. Regulated expression of hemoglobin and flavohemoglobin proteins postponed the onset of rpoS expression relative to plasmid bearing control cells. This change in expression pattern coincided with the expression pattern of stationary-phase specific genes including sigma(S)-dependent and sigma(S)-independent genes. Furthermore, several genes known to affect rpoS transcription, rpoS mRNA stability and sigma(S) turnover were regulated in such a manner as to ultimately lower the cellular level of sigma(S) in all globin-expressing strains. In a strain harboring an rpoS-lacZ fusion, lacZ expression correlated with acetate accumulation, a metabolite that is known to activate rpoS transcription, but not with growth. Therefore, we hypothesize that reduced excretion of acetate in globin expressing cells prevents induction of stationary phase specific genes. Additionally, several genes responding to carbon starvation (e.g. csrAB, cstA, sspA) were expressed at lower levels in globin-expressing cells. These findings are in good agreement with previous reports showing a more efficient energy household, i.e. also reduced glucose consumption, in hemoglobin- and flavohemoglobin-expressing cells relative to controls.
我们分析了表达天然和工程化细菌珠蛋白的大肠杆菌MG1655的基因表达,以确定在氧气受限条件下相对于对照细胞导致表型性状改善的分子机制。相对于携带质粒的对照细胞,血红蛋白和黄素血红蛋白蛋白的调控表达推迟了rpoS表达的起始。这种表达模式的变化与包括依赖sigma(S)和不依赖sigma(S)的基因在内的稳定期特异性基因的表达模式一致。此外,几个已知影响rpoS转录、rpoS mRNA稳定性和sigma(S)周转的基因以最终降低所有表达珠蛋白菌株中sigma(S)细胞水平的方式受到调控。在一个含有rpoS-lacZ融合的菌株中,lacZ表达与乙酸积累相关,乙酸是一种已知可激活rpoS转录的代谢物,但与生长无关。因此,我们假设表达珠蛋白的细胞中乙酸排泄减少可防止稳定期特异性基因的诱导。此外,几个对碳饥饿有反应的基因(如csrAB、cstA、sspA)在表达珠蛋白的细胞中表达水平较低。这些发现与先前的报道非常一致,即相对于对照,表达血红蛋白和黄素血红蛋白的细胞中能量利用更高效,即葡萄糖消耗也减少。