Yeast catalase A (Cta1p) contains two peroxisomal targeting signals (SSNSKF) localized at its C-terminus and within the N-terminal third of the protein, which both can target foreign proteins to peroxisomes. In the present study we demonstrated that Cta1p can also enter mitochondria, although the enzyme lacks a classical mitochondrial import sequence. Cta1p co-targeting was studied in a catalase A null mutant after growth on different carbon sources, and expression of a Cta1p–GFP (green fluorescent protein)-fusion protein or a Cta1p derivative containing either a c-Myc epitope (Cta1pmyc) or a SKF-extended tag (Cta1pmyc-SKF). Peroxisomal and mitochondrial co-import of catalase A were tested qualitatively by fluorescence microscopy and functional complementation of a Δcta1 null mutation, and quantitatively by subcellular fractionation followed by Western blot analysis and enzyme activity assays. Efficient Cta1p import into peroxisomes was observed when cells were cultivated under peroxisome-inducing conditions (i.e. growth on oleate), whereas significant co-import of Cta1p–GFP into mitochondria occurred when cells were grown under respiratory conditions that favour oxygen stress and ROS (reactive oxygen species) accumulation within this organelle. In particular, when cells were grown on the non-fermentable carbon source raffinose, respiration is maximally enhanced, and catalase A was efficiently targeted to the mitochondrial matrix where it presumably functions as scavenger of H2O2 and mitochondrial-derived ROS.
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Research Article|
June 01 2004
Dual targeting of yeast catalase A to peroxisomes and mitochondria
Ventsislava Y. PETROVA;
Ventsislava Y. PETROVA
1
*Angewandte Molekularbiologie, Universität des Saarlandes, D-66041 Saarbrücken, Germany
†Department of General and Industrial Microbiology, Faculty of Biology, St. Kliment Ohridski University, Sofia, Bulgaria
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Diane DRESCHER;
Diane DRESCHER
1
*Angewandte Molekularbiologie, Universität des Saarlandes, D-66041 Saarbrücken, Germany
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Anna V. KUJUMDZIEVA;
Anna V. KUJUMDZIEVA
†Department of General and Industrial Microbiology, Faculty of Biology, St. Kliment Ohridski University, Sofia, Bulgaria
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Manfred J. SCHMITT
Manfred J. SCHMITT
2
*Angewandte Molekularbiologie, Universität des Saarlandes, D-66041 Saarbrücken, Germany
2To whom correspondence should be addressed (e-mail mjs@microbiol.uni-sb.de).
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Publisher: Portland Press Ltd
Received:
January 07 2004
Revision Received:
March 02 2004
Accepted:
March 04 2004
Accepted Manuscript online:
March 04 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2004
2004
Biochem J (2004) 380 (2): 393–400.
Article history
Received:
January 07 2004
Revision Received:
March 02 2004
Accepted:
March 04 2004
Accepted Manuscript online:
March 04 2004
Citation
Ventsislava Y. PETROVA, Diane DRESCHER, Anna V. KUJUMDZIEVA, Manfred J. SCHMITT; Dual targeting of yeast catalase A to peroxisomes and mitochondria. Biochem J 1 June 2004; 380 (2): 393–400. doi: https://doi.org/10.1042/bj20040042
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