Identification of an alternative oxidase induction motif in the promoter region of the aod-1 gene in Neurospora crassa.
The nuclear aod-1 gene of Neurospora crassa encodes the alternative oxidase and is induced when the standard cytochrome-mediated respiratory chain of mitochondria is inhibited. To study elements of the pathway responsible for alternative oxidase induction, we generated a series of mutations in the region upstream from the aod-1 structural gene and transformed the constructs into an aod-1 mutant strain. Transformed conidia were plated on media containing antimycin A, which inhibits the cytochrome-mediated electron transport chain so that only cells expressing alternative oxidase will grow. Using this functional in vivo assay, we identified an alternative oxidase induction motif (AIM) that is required for efficient expression of aod-1. The AIM sequence consists of two CGG repeats separated by 7 bp and is similar to sequences known to be bound by members of the Zn(II)2Cys6 binuclear cluster family of transcription factors. The AIM motif appears to be conserved in other species found in the order Sordariales.
Chae
Michael S
MS
Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
Lin
Colin C
CC
Kessler
Katherine E
KE
Nargang
Cheryl E
CE
Tanton
Lesley L
LL
Hahn
Leanne B
LB
Nargang
Frank E
FE
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004579:Electron Transport; D045222:Electron Transport Chain Complex Proteins / Q000235:genetics / Q000378:metabolism; D004790:Enzyme Induction / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D009492:Neurospora crassa / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D010088:Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics*; D010940:Plant Proteins; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D014170:Transformation, Genetic; D000099188:Alternative Oxidase
D017931:DNA Primers; D004271:DNA, Fungal; D045222:Electron Transport Chain Complex Proteins; D005656:Fungal Proteins; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D000099188:Alternative Oxidase