Identification of the gene encoding the sole physiological fumarate reductase in Shewanella oneidensis MR-1. J Basic Microbiol 2003;43(4):312-27
Date
07/23/2003Pubmed ID
12872312DOI
10.1002/jobm.200390034Scopus ID
2-s2.0-0041927962 (requires institutional sign-in at Scopus site) 41 CitationsAbstract
Shewanella oneidensis MR-1 is a Gram-negative, nonfermentative rod with a complex electron transport system which facilitates its ability to use a variety of terminal electron acceptors, including fumarate, for anaerobic respiration. CMTn-3, a mutant isolated by transposon (TnphoA) mutagenesis, can no longer use fumarate as an electron acceptor; it lacks fumarate reductase activity as well as a 65-kDa soluble tetraheme flavocytochrome c. The sequence of the TnphoA-flanking genomic DNA of CMTn-3 did not align to those for fumarate reductase or related electron transport genes from other bacteria. Sequence analysis of the MR-1 genomic database demonstrated that an open reading frame encoding a 65-kDa tetraheme cytochrome c with sequence similarity to the fumarate reductase from S. frigidimarina NCIMB400 was found 8 kb away from the TnphoA-flanking genomic DNA of CMTn-3. PCR analysis demonstrated that a large deletion (>or=9.2 kb and <or=11 kb) of genomic DNA occurred in CMTn-3 as a result of TnphoA insertion. This deletion included at least half of the fumarate reductase gene as well as approximately 8 kb of upstream DNA. Complementation of CMTn-3 with the fumarate reductase gene plus 0.5-kb of upstream DNA restored growth on fumarate. These studies explicitly define the sole physiological fumarate reductase gene from the several possibilities suggested by the genomic sequence of MR-1. Surprisingly, the fumarate reductase gene plus 0.77-kb upstream DNA from S. frigidimarina NCIMB400 did not complement CMTn-3.
Author List
Maier TM, Myers JM, Myers CRMESH terms used to index this publication - Major topics in bold
DNA Transposable ElementsFumarates
Shewanella
Succinate Dehydrogenase
