The PmSOD1 gene of the protistan parasite Perkinsus marinus complements the sod2Delta mutant of Saccharomyces cerevisiae, and directs an iron superoxide dismutase to mitochondria.
The facultative intracellular oyster parasite, Perkinsus marinus, taxonomically related to both dinoflagellates and apicomplexan parasites, possesses at least two distinct genes (PmSOD1 and PmSOD2) predicted to encode iron-containing superoxide dismutases (Fe-SOD). The present study demonstrates that PmSOD1 complements a Saccharomyces cerevisiae mutant lacking the mitochondrial manganese-containing SOD (Mn-SOD), whereas PmSOD2 complements an Escherichia coli mutant lacking genes for cytosolic SOD activities. Mitochondria isolated from complemented yeast contain an SOD activity susceptible to inhibition by hydrogen peroxide, but resistant to cyanide, both characteristics of Fe-SODs. In cultured P. marinus trophozoites, indirect immunofluorescence using anti-PmSOD1 antibodies shows colocalization of PmSOD1 product with the mitochondrial marker MitoTracker Red. Further analysis of the leader sequence of the predicted PmSOD1 product revealed similarities to a mitochondrial targeting domain, an unusual observation for Fe-SODs, which are typically localized in the cytoplasm. These results suggest that PmSOD1 encodes a mitochondrial Fe-SOD, which may contribute to P. marinus resistance to exogenous oxidative damage in host phagocytes. The present study constitutes the first report of an endogenous Fe-SOD that is directed to the mitochondria, and suggests that mitochondria targeting sequences have been conserved among diverse branches of the eukaryotes, including the early protista. It also illustrates the potential of complementation-based approaches for further gene discovery and characterization in P. marinus.
Schott
Eric J
EJ
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 East Pratt Street, Baltimore 21202, USA.
Vasta
Gerardo R
GR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002244:Carbon / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D056890:Eukaryota / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D005816:Genetic Complementation Test; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008928:Mitochondria / Q000378:metabolism*; D008957:Models, Genetic; D009154:Mutation; D010269:Paraquat / Q000494:pharmacology; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology; D016415:Sequence Alignment; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology*; D000072105:Superoxide Dismutase-1; D014170:Transformation, Genetic; D000099156:Superoxide Dismutase 2
D002244:Carbon; D006861:Hydrogen Peroxide; D010269:Paraquat; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D000099156:Superoxide Dismutase 2; C000606290:SOD1 protein, human