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首页> 外文期刊>Journal of bacteriology >Dissimilatory Fe(III) and Mn(IV) Reduction by Shewanella putrefaciens Requires ferE, a Homolog of the pulE (gspE) Type II Protein Secretion Gene
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Dissimilatory Fe(III) and Mn(IV) Reduction by Shewanella putrefaciens Requires ferE, a Homolog of the pulE (gspE) Type II Protein Secretion Gene

机译：腐烂希瓦氏菌的异化铁（III）和锰（IV）还原需要ferE，这是pulE（gspE）II型蛋白质分泌基因的同源物。

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Shewanella putrefaciens strain 200 respires anaerobically on a wide range of compounds as the sole terminal electron acceptor, including ferric iron [Fe(III)] and manganese oxide [Mn(IV)]. Previous studies demonstrated that a 23.3-kb S. putrefaciens wild-type DNA fragment conferred metal reduction capability to a set of respiratory mutants with impaired Fe(III) and Mn(IV) reduction activities (T. DiChristina and E. DeLong, J. Bacteriol. 176:1468–1474, 1994). In the present study, the smallest complementing fragment was found to contain one open reading frame (ORF) (ferE) whose translated product displayed 87% sequence similarity to Aeromonas hydrophila ExeE, a member of the PulE (GspE) family of proteins found in type II protein secretion systems. Insertional mutants E726 and E912, constructed by targeted replacement of wild-type ferE with an insertionally inactivated ferE construct, were unable to respire anaerobically on Fe(III) or Mn(IV) yet retained the ability to grow on all other terminal electron acceptors. Nucleotide sequence analysis of regions flanking ferE revealed the presence of one partial and two complete ORFs whose translated products displayed 55 to 70% sequence similarity to the PulD, -F, and -G homologs of type II secretion systems. A contiguous cluster of 12 type II secretion genes (pulC to -N homologs) was found in the unannotated genome sequence of Shewanella oneidensis (formerly S. putrefaciens) MR-1. A 91-kDa heme-containing protein involved in Fe(III) reduction was present in the peripheral proteins loosely attached to the outside face of the outer membrane of the wild-type and complemented (Fer+) B31 transconjugates yet was missing from this location in Fer mutants E912 and B31 and in uncomplemented (Fer?) B31 transconjugates. Membrane fractionation studies with the wild-type strain supported this finding: the 91-kDa heme-containing protein was detected with the outer membrane fraction and not with the inner membrane or soluble fraction. These findings provide the first genetic evidence linking dissimilatory metal reduction to type II protein secretion and provide additional biochemical evidence supporting outer membrane localization of S. putrefaciens proteins involved in anaerobic respiration on Fe(III) and Mn(IV).
机译：腐烂希瓦氏菌菌株在作为唯一末端电子受体的多种化合物上厌氧呼吸，包括三价铁[Fe（III）]和氧化锰[Mn（IV）]。先前的研究表明，存在23.3kb的 S。腐烂的野生型DNA片段赋予一组具有减弱的Fe（III）和Mn（IV）还原活性的呼吸突变体金属还原能力（T. DiChristina和E. DeLong，J。Bacteriol。176：1468– 1474，1994）。在本研究中，发现最小的互补片段包含一个开放阅读框（ORF）（ ferE ），其翻译产物与 Aeromonas hydrophila ExeE的序列相似性为87％，在II型蛋白质分泌系统中发现的PulE（GspE）蛋白质家族的成员。通过用插入的灭活的 ferE 构建体有针对性地替换野生型 ferE 而构建的插入突变体E726和E912无法在Fe（III）或Mn（IV）上厌氧呼吸），但仍具有在所有其他末端电子受体上生长的能力。对 ferE 侧翼区域的核苷酸序列分析显示，存在一个部分和两个完整的ORF，其翻译产物与II型分泌系统的PulD，-F和-G同源物显示55-70％的序列相似性。在 Shewanella oneidensis 的未注释基因组序列中发现了12个II型分泌基因（ pulC 至- N 同源物）的连续簇（以前是< （em> S。putrefaciens ）MR-1。参与Fe（III）还原的含91 kDa血红素的蛋白质存在于与野生型外膜外表面松散连接且互补的（Fer + ）B31的周围蛋白质中Fer突变体E912和B31和非互补的（Fer ？）B31反轭合物中都没有从该位置找到反式轭合物。用野生型菌株进行膜分离研究支持了这一发现：用外膜部分而不是内膜或可溶性部分检测到含91 kDa血红素的蛋白质。这些发现提供了第一个将异化金属还原与II型蛋白质分泌联系起来的遗传证据，并提供了其他生物化学证据来支持 S的外膜定位。腐乳蛋白参与Fe（III）和Mn（IV）的厌氧呼吸。

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