Isolation, Characterization, and Expression of the Gene Encoding the b 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 b subunit of the Blastocladiella mitochondrial processing peptidase (b-MPP). The predicted b-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 b-MPP mRNA levels decrease significantly during B. emersonii sporulation, reaching basal levels in the zoospore stage. The amount of b-MPP protein, determined in Western blots, unlike its mRNA, does not vary significantly throughout the fungal life cycle.