The Journal of Biochemistry
Online ISSN : 1756-2651
Print ISSN : 0021-924X
Identification of a Cryptic N-Terminal Signal in Saccharomyces cerevisiae Peroxisomal Citrate Synthase That Functions in Both Peroxisomal and Mitochondrial Targeting
Jeong Goo LeeSeong Pil ChoHeon Sik LeeCheong Ho LeeKyung Sook BaePil Jae Maeng
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JOURNAL FREE ACCESS

2000 Volume 128 Issue 6 Pages 1059-1072

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Abstract

Saccharomyces cerevisiae has three distinct citrate synthases, two located in mitochondria (mature Citlp and Cit3p) and one in peroxisomes (mature Cit2p). While the precursor of the major mitochondrial enzyme, Citlp, has a signal for mitochondrial targeting at its N-terminus (MTS), Cit2p has one for peroxisomal targeting (PTS1) at its C-terminus. We have previously shown that the N-terminal segment of Cit2p is removed during import into peroxisomes [Lee, H. S. et al. (1994) Kor. J. Microbiol. 32, 558-564], which implied the presence of an additional N-terminal sorting signal. To analyze the function of the N-terminal region of Cit2p in protein trafficking, we constructed the N-terminal domain-swapped versions of Citlp and Cit2p. Both fusions, Cit1::Cit2 and Cit2::Citl, complemented the glutamate auxotrophy caused by the double-disruption of the CIT1 and CIT2 genes. In addition, part of the Cit2::Cit1 fusion protein, as well as Cit1::Cit2, was shown to be transported into both mitochondria and peroxisomes. The subcellular localization of the recombinant fusion proteins containing various N-terminal segments of Cit2p fused to a mutant version of green fluorescent protein (GFP2) was also examined. As a result, we found that the 20-amino acid N-terminal segment of Cit2p contains a cryptic cleavable targeting signal for both peroxisomes and mitochondria. In addition, we show that the peroxisomal import process mediated by the N-terminal segment of Cit2p was not affected by the disruption of either PEX5 (encoding PTS1 receptor) or PEX7 (encoding PTS2 receptor).

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© The Japanese Biochemical Society
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