Characterization and submitochondrial localization of the alpha subunit of the mitochondrial processing peptidase from the aquatic fungus Blastocladiella emersonii.
In an effort to investigate the molecular mechanisms responsible for the drastic morphological changes the mitochondria go through during the life cycle of the aquatic fungus Blastocladiella emersonii, the gene encoding the alpha subunit of the mitochondrial processing peptidase (alpha-MPP) was isolated. Nucleotide sequence analysis revealed that the predicted alpha-MPP polypeptide comprises 474 amino acids with a calculated molecular mass of 51,900 Da, presenting a characteristic mitochondrial signal sequence. Northern blot analysis indicated a single 1.4-kb transcript encoding the B. emersonii alpha-MPP, whose levels decrease during sporulation, becoming very low in the zoospore, and increase again during germination. Despite these variations in mRNA concentration, B. emersonii alpha-MPP protein levels do not change significantly during the life cycle of the fungus, as observed in Western blots. Experiments to investigate the submitochondrial localization of B. emersonii alpha-MPP and beta-MPP were also carried out, and the results indicated that both subunits are associated with the mitochondrial inner membrane, possibly as part of the bc1 complex, as described for plants.
Rocha
C R
CR
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo 05599-970, Brazil.
Gomes
S L
SL
D000595:Amino Acid Sequence; D001483:Base Sequence; D001754:Blastocladiella / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D015153:Blotting, Western; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D007425:Intracellular Membranes / Q000201:enzymology; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013367:Submitochondrial Particles / Q000201:enzymology; D014158:Transcription, Genetic; D000096122:Mitochondrial Processing Peptidase
D004271:DNA, Fungal; D008666:Metalloendopeptidases; D000096122:Mitochondrial Processing Peptidase