Disruption of the gene encoding the 78-kilodalton subunit of the peripheral arm of complex I in Neurospora crassa by repeat induced point mutation (RIP).
We have used the procedure of sheltered RIP to generate mutants of the 78-kDa protein of the peripheral arm of Neurospora crassa complex I. The nuclei containing the mutations were initially isolated as one component of a heterokaryon but subsequent analysis showed that nuclei containing null alleles of the gene could be propagated as homokaryons. This demonstrates that the gene does not serve an essential function. Sequence analysis of one allele shows that 61 transition mutations were created resulting in 39 amino-acid changes including the introduction of four stop codons. Mutant strains grow at a slower rate than wild-type and exhibit a decrease in the production of conidia. Electron paramagnetic spectroscopy of mutant mitochondria suggest that they are deficient in Fe-S clusters N-1, N-3, and N-4.
Harkness
T A
TA
Department of Biological Sciences, University of Alberta, Edmonton, Canada.
Rothery
R A
RA
Weiner
J H
JH
Werner
S
S
Azevedo
J E
JE
Videira
A
A
Nargang
F E
FE
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003433:Crosses, Genetic; D003580:Cytochromes / Q000737:chemistry; D004271:DNA, Fungal / Q000235:genetics*; D004578:Electron Spin Resonance Spectroscopy; D042967:Electron Transport Complex I; D008040:Genetic Linkage; D007506:Iron-Sulfur Proteins / Q000235:genetics* / Q000502:physiology; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009492:Neurospora crassa / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D017354:Point Mutation; D014170:Transformation, Genetic
D003580:Cytochromes; D004271:DNA, Fungal; D007506:Iron-Sulfur Proteins; D009247:NADH, NADPH Oxidoreductases; D042967:Electron Transport Complex I