A single precursor protein for ferrochelatase-I from Arabidopsis is imported in vitro into both chloroplasts and mitochondria.
Ferrochelatase is the last enzyme of heme biosynthesis and in higher plants is found in both chloroplasts and mitochondria. We have isolated cDNAs for two isoforms of ferrochelatase from Arabidopsis thaliana, both of which are imported into isolated chloroplasts. In this paper we show that ferrochelatase-I is also imported into isolated pea mitochondria with approximately the same efficiency as into chloroplasts. Processing of the precursor was observed with both chloroplast stroma and mitochondrial matrix extracts. This was inhibited by EDTA, indicating it was due to the specific processing proteases. The specificity of import was verified by the fact that the mitochondrial preparation did not import the precursor of the light-harvesting chlorophyll a/b protein precursor or the precursor of porphobilinogen deaminase, an earlier enzyme of tetrapyrrole biosynthesis, both of which are exclusively chloroplast-located. Furthermore, import of ferrochelatase-I precursor into mitochondria was inhibited by valinomycin, but this had no effect on its import into chloroplasts. Thus a single precursor molecule is recognized by the import machinery of the two organelles. The implications for the targeting of ferrochelatase in a possible protective role against photooxidative stress are discussed.
Chow
K S
KS
Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge CB2 3EA, United Kingdom.
Singh
D P
DP
Roper
J M
JM
Smith
A G
AG
D017360:Arabidopsis / Q000378:metabolism*; D001692:Biological Transport; D002736:Chloroplasts / Q000378:metabolism*; D018076:DNA, Complementary; D005294:Ferrochelatase / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D018532:Pisum sativum / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
D018076:DNA, Complementary; D011498:Protein Precursors; D005294:Ferrochelatase