Abstract
Carnitine acetyltransferase (CAT) is present in mitochondria and peroxisomes of oleate-grown Saccharomyces cerevisiae. Both proteins are encoded by the same gene, YCAT, which encodes a protein with a mitochondrial targeting signal (MTS) at the N-terminus, and a peroxisomal targeting signal type 1 (PTS-1) at the C-terminus. Deletion of both motifs revealed the presence of an additional internal targeting sequence. Import of CAT via this internal signal was shown to be dependent on PAS10, a protein which is required for the import of PTS-1 containing proteins. An interaction of PAS10 with this internal targeting signal was demonstrated using the yeast two-hybrid technique. Expression of the YCAT gene behind a heterologous promoter resulted in loss of peroxisomal targeting, indicating that differential targeting is controlled at transcriptional or translational level. Determination of the 5'-ends of YCAT mRNAs revealed that YCAT transcripts initiating after the first AUG were present in oleate-grown cells. These transcripts were virtually absent in acetate- or glycerol-grown cells. We propose that in response to oleate, shorter transcripts are produced from which the peroxisomal form of CAT is translated, resulting in a CAT protein without a MTS, which can be targeted to peroxisomes.
MeSH Terms
Base Sequence
Carnitine O-Acetyltransferase/genetics,metabolism
DNA Primers
Gene Expression Regulation, Fungal
Genes, Fungal
Microbodies/enzymology
Mitochondria/enzymology
Molecular Sequence Data
Oleic Acid
Oleic Acids/metabolism
Protein Sorting Signals/genetics,metabolism
Saccharomyces cerevisiae/enzymology,genetics
Signal Transduction
Transcription, Genetic
Chemicals
DNA Primers
Oleic Acids
Protein Sorting Signals
Oleic Acid
Carnitine O-Acetyltransferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Elgersma Y
Department of Biochemistry, E.C. Slater Institute, Amsterdam, The Netherlands.
van Roermund C W
Wanders R J
Tabak H F
References (27)
27 references, click to expand
-
Fatty acid beta-oxidation system in microbodies of n-alkane-grown Candida tropicalis.
Eur J Biochem. 1978 Feb;83(2):609-13
PMID: 204485
-
The membrane of peroxisomes in Saccharomyces cerevisiae is impermeable to NAD(H) and acetyl-CoA under in vivo conditions.
EMBO J. 1995 Jul 17;14(14):3480-6
PMID: 7628449
-
Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase.
Cell. 1982 Jan;28(1):145-54
PMID: 7039847
-
Peroxisomal and mitochondrial carnitine acetyltransferases in alkane-grown yeast Candida tropicalis.
Eur J Biochem. 1982 May;124(1):205-10
PMID: 7084226
-
Purification and properties of carnitine acetyltransferase from rat liver.
J Biochem. 1983 Feb;93(2):439-51
PMID: 6404901
-
Biosynthesis and turnover of carnitine acetyltransferase in rat liver.
J Biochem. 1983 Feb;93(2):453-9
PMID: 6841345
-
Characterization of peroxisomal and mitochondrial carnitine acetyltransferases purified from alkane-grown Candida tropicalis.
Eur J Biochem. 1984 Feb 1;138(3):445-9
PMID: 6692827
-
Synthesis in vitro of precursor-type carnitine acetyltransferase with messenger RNA from Candida tropicalis.
Eur J Biochem. 1984 Feb 1;138(3):451-7
PMID: 6692828
-
Acyl-CoA oxidase contains two targeting sequences each of which can mediate protein import into peroxisomes.
EMBO J. 1988 Apr;7(4):1167-73
PMID: 3402435
-
Carnitine.
Annu Rev Biochem. 1988;57:261-83
PMID: 3052273
-
Comparative enzymology of beta-oxidation.
Biochem Soc Trans. 1988 Jun;16(3):418-20
PMID: 3053289
-
A conserved tripeptide sorts proteins to peroxisomes.
J Cell Biol. 1989 May;108(5):1657-64
PMID: 2654139
-
A novel genetic system to detect protein-protein interactions.
Nature. 1989 Jul 20;340(6230):245-6
PMID: 2547163
-
Generation from a single gene of two mRNAs that encode the mitochondrial and peroxisomal serine:pyruvate aminotransferase of rat liver.
J Biol Chem. 1990 May 5;265(13):7513-9
PMID: 2332438
-
Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency: identification of a new inborn error of mitochondrial fatty acid beta-oxidation.
J Inherit Metab Dis. 1990;13(3):311-4
PMID: 2122092
-
Targeting efficiencies of various permutations of the consensus C-terminal tripeptide peroxisomal targeting signal.
FEBS Lett. 1992 Jun 29;305(2):133-6
PMID: 1618341
-
Molecular evolution of alanine/glyoxylate aminotransferase 1 intracellular targeting. Analysis of the marmoset and rabbit genes.
Eur J Biochem. 1992 Jul 15;207(2):757-66
PMID: 1339350
-
Isolation of peroxisome assembly mutants from Saccharomyces cerevisiae with different morphologies using a novel positive selection procedure.
J Cell Biol. 1992 Oct;119(1):153-62
PMID: 1356111
-
Cloning and sequencing of a cDNA encoding Saccharomyces cerevisiae carnitine acetyltransferase. Use of the cDNA in gene disruption studies.
J Biol Chem. 1993 Jan 25;268(3):1824-9
PMID: 8420957
-
Two independent peroxisomal targeting signals in catalase A of Saccharomyces cerevisiae.
J Cell Biol. 1993 Feb;120(3):665-73
PMID: 8425895
-
Presence of carnitine acetyltransferase in peroxisomes and in mitochondria of oleic acid-grown Saccharomyces cerevisiae.
FEMS Microbiol Lett. 1993 Aug 15;112(1):31-4
PMID: 8405946
-
Assay of succinate dehydrogenase activity by a colorimetric-continuous method using iodonitrotetrazolium chloride as electron acceptor.
Anal Biochem. 1993 Aug 1;212(2):506-9
PMID: 8214593
-
The ethanol-inducible YAT1 gene from yeast encodes a presumptive mitochondrial outer carnitine acetyltransferase.
J Biol Chem. 1993 Dec 25;268(36):27428-39
PMID: 8262985
-
PAS10 is a tetratricopeptide-repeat protein that is essential for the import of most matrix proteins into peroxisomes of Saccharomyces cerevisiae.
Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11782-6
PMID: 8265627
-
An efficient positive selection procedure for the isolation of peroxisomal import and peroxisome assembly mutants of Saccharomyces cerevisiae.
Genetics. 1993 Nov;135(3):731-40
PMID: 7507454
-
PAS7 encodes a novel yeast member of the WD-40 protein family essential for import of 3-oxoacyl-CoA thiolase, a PTS2-containing protein, into peroxisomes.
EMBO J. 1994 Oct 17;13(20):4908-18
PMID: 7957058
-
Localization of carnitine acetyltransferase in peroxisomes and in mitochondria of n-alkane-grown Candida tropicalis.
FEBS Lett. 1978 Dec 1;96(1):37-40
PMID: 215452