A temperature-sensitive mutant of Neurospora crassa deficient in cytochrome b.

Abstract

A nuclear gene mutant of Neurospora crassa designated cyb-3 is deficient in cytochrome b and coenzyme QH2-cytochrome c reductase. Nearly normal when grown at 25 degrees C, the strain expresses a mutant phenotype at 38 degrees C. Mitochondria from cyb-3 mycelium, which has undergone 3-4 mass doublings at the elevated temperature, possess 3-fold less cytochrome b, 2-fold more cytochrome, c, 5-fold less coenzyme QH2-cytochrome c reductase activity, and require 3-fold less antimycin A per milligram of protein to inhibit NADH oxidation that do wild type mitochondria. The activity of coenzyme QH2-cytochrome c reductase declines rather slowly in cultures of cyb-3 transferred to 38 degrees C, and the in vitro thermostability of the enzyme is very similar in wild type and mutant mitochondria. Therefore, the mutation may decrease synthesis of impair integration into the membrane of cytochrome b and perhaps other proteins of the enzyme comple.