GENETIC-ANALYSIS OF THE CYTOCHROME-C-AA3 BRANCH OF THE BRADYRHIZOBIUM-JAPONICUM RESPIRATORY-CHAIN

被引:86
作者
BOTT, M [1 ]
BOLLIGER, M [1 ]
HENNECKE, H [1 ]
机构
[1] SWISS FED INST TECHNOL,INST MIKROBIOL,SCHMELZBERGSTR 7,CH-8092 ZURICH,SWITZERLAND
关键词
D O I
10.1111/j.1365-2958.1990.tb00576.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Further genetic evidence is provided here that Brady-rhizobium japonicum possesses a mitochondria-like electron-transport pathway: 2[H]-->UQ-->bc1-->c-->aa3 -->O2. Two Tn5-induced mutants, COX122 and COX132, having cytochrome c oxidase-negative phenotypes, were obtained and characterized. Mutant COX122 was defective in a novel gene, named cycM, which was responsible for the synthesis of a c-type cytochrome with an M(r) of 20 000 (20K). This 20K cytochrome c appeared to catalyse electron transport from the cytochrome bc1 complex to the aa3-type terminal oxidase and, unlike mitochondrial cytochrome c, was membrane-bound in B. japonicum. The Tn5 insertion of mutant COX132 was localized in coxA, the structural gene for subunit 1 of cytochrome aa3. This finding also led to the cloning and sequencing of the corresponding wild-type coxA gene that encoded a 541-amino-acid protein with a predicted M(r) of 59247. The CoxA protein shared about 60% sequence identity with the cytochrome aa3 subunit l of mitochondria. The B. japonicum cycM and coxA mutants were able to fix nitrogen in symbiosis with soybean (Fix + ). In contrast, mutants described previously which lacked the bc1 complex did not develop into endosymbiotic bacteroids and were thus Fix -. The data suggest that a symbiosis-specific respiratory chain exists in B. japonicum in which the electrons branch off at the bc1 complex.
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页码:2147 / 2157
页数:11
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