Cao Dongmei, Ji Wenhui, Fu Qiang, Lu Chenping, Wang Hengan, Sun Jianhe, Yan Yaxian
Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China;
Key Laboratory of Animal Disease Diagnosis and Immunology of Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China.
FEMS Microbiol Lett. 2015 Oct;362(19). doi: 10.1093/femsle/fnv149. Epub 2015 Sep 3.
It has been earlier hypothesized that lysogenic infection with Stx-encoding phages influences protein expression in the bacterial host, and therefore, some differentially expressed proteins could affect survival characteristics and pathogenicity. We compared the protein expression profiles of the host MG1655 and lysogens by 2D electrophoresis. Four different genes identified were all related to Fe/S subunit production, namely, nfuA, fdoH, sdhB and ftnA. To explore the role of nfuA in the biology of Stx prophage lysogeny, gene knockout experiments and phage lysogenic conversion were performed. The inactivation of nfuA caused the prophage to enter its lytic life cycle, especially under an iron-depleted condition. A similar activity was also detected in the Escherichia coli O157:H7 strain from which the Stx phage Min 27 was originally isolated. NfuA might be the positive regulator of genes controlling lysogenic cycle such as cI, cII and cIII since their transcriptional level was significantly reduced in nfuA deletion mutant as shown by qRT-PCR. We conclude that NfuA is essential for maintenance of Stx phage lysogeny in host's genetic reservoir under iron-deficient condition.
早期曾有假设认为,携带stx编码噬菌体的溶原性感染会影响细菌宿主中的蛋白质表达,因此,一些差异表达的蛋白质可能会影响生存特征和致病性。我们通过二维电泳比较了宿主MG1655和溶原菌的蛋白质表达谱。鉴定出的四个不同基因均与铁硫亚基的产生有关,即nfuA、fdoH、sdhB和ftnA。为了探究nfuA在stx原噬菌体溶原性生物学中的作用,进行了基因敲除实验和噬菌体溶原性转化。nfuA的失活导致原噬菌体进入其裂解生命周期,尤其是在缺铁条件下。在最初分离出stx噬菌体Min 27的大肠杆菌O157:H7菌株中也检测到了类似的活性。nfuA可能是控制溶原周期的基因如cI、cII和cIII的正调控因子,因为qRT-PCR显示,在nfuA缺失突变体中它们的转录水平显著降低。我们得出结论,在缺铁条件下,NfuA对于在宿主基因库中维持stx噬菌体溶原性至关重要。