Genes Linking Copper Trafficking and Homeostasis to the Biogenesis and Activity of the cbb (3)-Type Cytochrome c Oxidase in the Enteric Pathogen Campylobacter jejuni.
Genes Linking Copper Trafficking and Homeostasis to the Biogenesis and Activity of the cbb (3)-Type Cytochrome c Oxidase in the Enteric Pathogen Campylobacter jejuni.
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DOI：
10.3389/fmicb.2021.683260
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发表时间：
2021
影响因子：
5.2
通讯作者：
Kelly DJ
中科院分区：
文献类型：
--
作者：
Garg N;Taylor AJ;Pastorelli F;Flannery SE;Jackson PJ;Johnson MP;Kelly DJ
Cbb3家族中的细菌C型血红铜氧化酶广泛存在于微需氧微生物中，这些微生物利用其高的氧结合亲和力在微氧生态位中生长。在嗜氧微创病原体中，C型氧化酶可能是感染所必需的，但与模型细菌相比，人们对它们的生物发生知之甚少。在这里，我们已经确定了与CBB3-氧化酶(CCO)组装和活性有关的基因，这是革兰氏阴性病原体空肠弯曲杆菌的最常见原因，它是人类食源性细菌性胃肠炎的最常见原因。CcoNOQP下游的一些未知功能基因对CCO活性是必需的(cj1483c和cj1486c)或重要的(cj1484c和cj1485c)；Cj1483是CcoH同源物，但Cj1484(命名为CcoZ)在结构上与MSMEG_4692相似，参与耻垢分枝杆菌QCR-氧化酶超复合体的形成。蓝色天然聚丙烯酰胺凝胶电泳法显示洗涤剂增溶膜有三条主带，其中一条带含有CcoZ、QCR和氧化酶亚基。推测的铜运输基因ccoI(Cj1155c)和ccos(Cj1154c)的缺失可使CCO活性部分恢复，而编码CCOG同源物的cj0369c的失活导致CCO活性的部分降低。缺失编码PCuAC(Cj0909)和Sco(Cj0911)周质铜伴侣同系物的操纵子可降低CCO活性，外源铜可部分恢复cj0911突变体的CCO活性。对编码细胞内金属动态平衡的几个基因的cj1161c-1166c基因缺失的表型分析表明，CopA(Cj1161c)或CopZ(Cj1162c)的失活导致细胞内铜升高和CCO活性降低，这种影响在外部高铜时加剧。因此，我们的工作确定了(I)额外的CCO亚基，(Ii)一组以前未描述的连接铜运输和CCO活性的基因，以及(Iii)在这种重要病原体中与铜稳态有关的联系。
Bacterial C-type haem-copper oxidases in the cbb3 family are widespread in microaerophiles, which exploit their high oxygen-binding affinity for growth in microoxic niches. In microaerophilic pathogens, C-type oxidases can be essential for infection, yet little is known about their biogenesis compared to model bacteria. Here, we have identified genes involved in cbb3-oxidase (Cco) assembly and activity in the Gram-negative pathogen Campylobacter jejuni, the commonest cause of human food-borne bacterial gastroenteritis. Several genes of unknown function downstream of the oxidase structural genes ccoNOQP were shown to be essential (cj1483c and cj1486c) or important (cj1484c and cj1485c) for Cco activity; Cj1483 is a CcoH homologue, but Cj1484 (designated CcoZ) has structural similarity to MSMEG_4692, involved in Qcr-oxidase supercomplex formation in Mycobacterium smegmatis. Blue-native polyacrylamide gel electrophoresis of detergent solubilised membranes revealed three major bands, one of which contained CcoZ along with Qcr and oxidase subunits. Deletion of putative copper trafficking genes ccoI (cj1155c) and ccoS (cj1154c) abolished Cco activity, which was partially restored by addition of copper during growth, while inactivation of cj0369c encoding a CcoG homologue led to a partial reduction in Cco activity. Deletion of an operon encoding PCuAC (Cj0909) and Sco (Cj0911) periplasmic copper chaperone homologues reduced Cco activity, which was partially restored in the cj0911 mutant by exogenous copper. Phenotypic analyses of gene deletions in the cj1161c–1166c cluster, encoding several genes involved in intracellular metal homeostasis, showed that inactivation of copA (cj1161c), or copZ (cj1162c) led to both elevated intracellular Cu and reduced Cco activity, effects exacerbated at high external Cu. Our work has therefore identified (i) additional Cco subunits, (ii) a previously uncharacterized set of genes linking copper trafficking and Cco activity, and (iii) connections with Cu homeostasis in this important pathogen.
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影响因子：
4.8
作者：
Horng, YC;Cobine, PA;Winge, DR
通讯作者：
Winge, DR
影响因子：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
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Sternberg MJ
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Kassem, Issmat I.;Candelero-Rueda, Rosario A.;Rajashekara, Gireesh
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Rajashekara, Gireesh