Characterization and Submitochondrial Localization of the α Subunit of the Mitochondrial Processing Peptidase from the Aquatic Fungus Blastocladiella emersonii

Abstract

ABSTRACT 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 α subunit of the mitochondrial processing peptidase (α-MPP) was isolated. Nucleotide sequence analysis revealed that the predicted α-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 α-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 α-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 α-MPP and β-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.