A cytosolic phospholipase A2-like protein in the filamentous fungus Aspergillus oryzae localizes to the intramembrane space of the mitochondria.
In mammalian cells, cytosolic phospholipase A(2) (cPLA(2)) displays a variety of activities through the release of arachidonic acid in response to cellular stimuli. In this study, we characterize the putative cPLA(2)-like protein, AoPlaA, in the filamentous fungus Aspergillus oryzae. When AoPlaA-EGFP was expressed in A. oryzae, it localized to the tubular structures that was costained by the marker dye for the mitochondria. A biochemical fractionation experiment showed that AoPlaA was present in the mitochondria-enriched fraction. The presence of an N-terminal cleavable mitochondrial targeting signal in AoPlaA was demonstrated by N-terminal amino acid sequence analysis, and by showing that chimeric proteins consisting of N-terminal 65 or 50 amino acids of AoPlaA fused to enhanced green fluorescent protein (EGFP) localized to the mitochondria. Submitochondrial fractionation of AoPlaA expressed in Saccharomyces cerevisiae demonstrated that AoPlaA localizes to the intramembrane space of the mitochondria. Taken together, the results presented here demonstrate a novel, submitochondrial localization of the cPLA(2)-like protein in the filamentous fungi.
Takaya
Kohei
K
Department of Biotechnology, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Higuchi
Yujiro
Y
Kitamoto
Katsuhiko
K
Arioka
Manabu
M
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology*; D005656:Fungal Proteins / Q000032:analysis* / Q000235:genetics; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008928:Mitochondria / Q000737:chemistry*; D051336:Mitochondrial Membranes / Q000737:chemistry*; D024101:Mitochondrial Proteins / Q000032:analysis* / Q000235:genetics; D008969:Molecular Sequence Data; D054467:Phospholipases A2 / Q000032:analysis* / Q000235:genetics; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
D005656:Fungal Proteins; D024101:Mitochondrial Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D054467:Phospholipases A2