Isolation, characterization, and expression of the gene encoding the beta subunit of the mitochondrial processing peptidase from Blastocladiella emersonii.
A 2.3-kb BamHI-KpnI fragment was isolated from a partial genomic library and shown by nucleotide sequence analysis to contain the entire coding region of the gene encoding the beta subunit of the Blastocladiella mitochondrial processing peptidase (beta-MPP). The predicted beta-MPP protein has 465 amino acids and a calculated molecular mass of 50.8 kDa. S1 nuclease protection assays revealed an intron, 209 bp in size, interrupting the coding region between the putative signal sequence and the mature protein. Northern blot analysis showed that beta-MPP mRNA levels decrease significantly during B. emersonii sporulation, reaching basal levels in the zoospore stage. The amount of beta-MPP protein, determined in Western blots, unlike its mRNA, does not vary significantly throughout the fungal life cycle.
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo 05599-970, Brazil.
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Costa Rocha
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Lopes Gomes
D000595:Amino Acid Sequence; D001483:Base Sequence; D001754:Blastocladiella / Q000201:enzymology* / Q000235:genetics*; D005800:Genes, Fungal; D015698:Genomic Library; D046911:Macromolecular Substances; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014158:Transcription, Genetic; D000096122:Mitochondrial Processing Peptidase
D046911:Macromolecular Substances; D008666:Metalloendopeptidases; D000096122:Mitochondrial Processing Peptidase