Reconstitution of succinate dehydrogenase in Bacillus subtilis by protoplast fusion.
Bacillus subtilis succinate dehydrogenase (SDH) is composed of two unequal subunits designated Fp (Mr, 65,000) and Ip (Mr. 28,000). The enzyme is structurally and functionally complexed to cytochrome b 558 (Mr, 19,000) in the membrane. A total of 21 B. subtilis SDH-negative mutants were isolated. The mutants fall into five phenotypic classes with respect to the presence and localization of the subunits of the SDH-cytochrome b558 complex. One class contains mutants with an inactive membrane-bound complex. Membrane-bound enzymatically active SDH could be reconstituted in fused protoplasts of selected pairs of SDH-negative mutants. Most likely reconstitution is due to the assembly of preformed subunits in the fused cells. On the basis of the reconstitution data, the mutants tested could be divided into three complementation groups. The combined data of the present and previous work indicate that the complementation groups correspond to the structural genes for the three subunits of the membrane-bound SDH-cytochrome b558 complex. A total of 31 SDH-negative mutants of B. subtilis have now been characterized. The respective mutations all map in the citF locus at 255 degrees on the B. subtilis chromosomal map. In the present paper, we have revised the nomenclature for the genetics of SDH in B. subtilis. All mutations which give an SDH-negative phenotype will be called sdh followed by an isolation number. The designation citF will be omitted, and the citF locus will be divided into three genes: sdhA, sdhB, and sdhC. Mutations in sdhA affect cytochrome b558, mutations in sdhB affect Fp, and mutations in sdhC affect Ip.
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001431:Bacteriological Techniques; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008561:Membrane Fusion; D009154:Mutation; D011523:Protoplasts / Q000201:enzymology*; D013385:Succinate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D009626:Terminology as Topic
D013385:Succinate Dehydrogenase