Characterization of mutations in the mitochondrial cytochrome b gene of Saccharomyces cerevisiae affecting the quinone reductase site (QN).

Abstract

The revertant [G33A]cytochrome b recently isolated from the [G33D]cytochrome b mutant [Coppée, J. Y., Tokutake, N., Marc, D., di Rago, J.-P., Miyoshi, H. & Colson, A.-M. (1994) FEBS Lett. 339, 1-6] exhibits cross resistance to center-N inhibitors 2-heptyl-4-hydroxyquinoline N-oxide (HQNO) and funiculosin and a spectral shift in the cytochrome b562 heme. This indicates that the conserved G33 residue is in the vicinity of this heme, and thus agrees with the previous suggestion that glycine may play a role in the helix packing around the hemes. The [S206L]cytochrome b and [M221K]cytochrome b respiratory-growth-deficient mutants [Lemesle-Meunier, D., Brivet-Chevillotte, P., di Rago, J. P., Slonimski, P. P., Bruel, C., Tron, T. & Forget, N. (1993) J. Biol. Chem. 268, 15,626-15,632], which synthesize cytochrome b and retain little or no bc1 complex activity, show no change in the reduction kinetics of cytochrome b via center P, which suggests that the oxidizing site is functional. Impairment of both the reduction and oxidation of heme b562 at the ubiquinone reduction center of the mitochondrial ubiquinone-cytochrome-c oxidoreductase site is, therefore, responsible for the deficient catalytic activity and respiratory growth in these strains.