Intragenic suppressors reveal long distance interactions between inactivating and reactivating amino acid replacements generating three-dimensional constraints in the structure of mitochondrial cytochrome b.
Revertants of nonfunctional cytochrome b mutants were isolated and characterized to determine how specific deleterious mutations in cytochrome b can be suppressed by secondary mutations not restoring a wild type protein. It was recently shown that the cytochrome b function can be recovered following various pseudo-wild type reversions at the level of the original site mutation or adjacent positions (di Rago, J.-P., Netter, P., and Slonimski, P. P. (1990) J. Biol. Chem. 265, 3332-3339). In the present study, we describe how the cytochrome b function can be recovered by secondary mutations in positions which are removed from the original mutation by up to more than 100 amino acids. Such revertant mutants are useful for the study of the three-dimensional structure of cytochrome b. The results of the analysis of four deficient mutations which affect a short region of the protein (positions 131-138 of the polypeptide chain) lead us to propose a possible mode of interactive combination between the first five putative transmembrane segments of cytochrome b within the membrane.
di Rago
J P
JP
Centre de Génétique Moléculaire, l'Université Pierre et Marie Curie, Gif-sur-Yvette, France.
Netter
P
P
Slonimski
P P
PP
D000595:Amino Acid Sequence; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D003573:Cytochrome b Group / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure; D007438:Introns; D046911:Macromolecular Substances; D008928:Mitochondria / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013489:Suppression, Genetic
D003062:Codon; D003573:Cytochrome b Group; D046911:Macromolecular Substances