Four mutants of AM1 which were deficient in the production of soluble -type cytochromes have been isolated and characterized. They were unable to grow on methanol, ethanol or methylamine and -type cytochromes were not detected in cell extracts by spectroscopy or in haem-stained SDS-polyacrylamide gels. Western blotting demonstrated that the mutants did not contain either the apo- or the holo-forms of cytochrome , although trace amounts of holo-cytochrome were detected. The mutants had only trace levels of methylamine dehydrogenase activity, suggesting a role for at least one of the -type cytochromes in the production of the active form of this enzyme. Two regions of the AM1 chromosome which encode genes required for the synthesis of -type cytochromes were identified and we have shown that, although the four mutants were phenotypically identical, they represent at least three complementation groups.


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