Cloning and identification of HEM14, the yeast gene for mitochondrial protoporphyrinogen oxidase.
A respiratory-defective mutant (C54) of Saccharomyces cerevisiae was found to have a phenotype consistent with a mutation in either mitochondrial protoporphyrinogen oxidase or ferrochelatase. The mutant is grossly deficient in hemes, accumulates protoporphyrin and is rescued by exogenous heme. The increased levels of protoporphyrin at the expense of heme is indicative of a block in one of the two last steps of the heme biosynthetic pathway. Complementation of C54 by a known ferrochelatase mutant suggested that the defect was most likely in HEM14 encoding protoporphyrinogen oxidase. A plasmid capable of complementing C54 was obtained by transformation with a yeast genomic plasmid library. A partial sequence of the insert identified the gene as reading frame YER014 of yeast chromosome V (GenBank Accession Number U18778). This reading frame codes for a protein homologous to human protoporphyrinogen oxidase. Disruption of this gene elicits a respiratory defect and accumulation of protoporphyrin. The phenotype of the null mutant together with the homology of YER014p to human protoporphyrinogen oxidase provide compelling evidence that YER014 is HEM14.
Glerum
D M
DM
Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
Shtanko
A
A
Tzagoloff
A
A
Gorman
N
N
Sinclair
P R
PR
D003001:Cloning, Molecular; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006418:Heme / Q000032:analysis; D008928:Mitochondria / Q000201:enzymology*; D024101:Mitochondrial Proteins; D010088:Oxidoreductases / Q000235:genetics*; D044925:Oxidoreductases Acting on CH-CH Group Donors; D010101:Oxygen Consumption / Q000235:genetics; D010641:Phenotype; D011166:Porphyrins / Q000032:analysis; D050768:Protoporphyrinogen Oxidase; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017385:Sequence Homology; D013045:Species Specificity
D024101:Mitochondrial Proteins; D011166:Porphyrins; D029701:Saccharomyces cerevisiae Proteins; D006418:Heme; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C493976:HEM14 protein, S cerevisiae; D050768:Protoporphyrinogen Oxidase