Elgersma Y, van Roermund C W, Wanders R J, Tabak H F
Department of Biochemistry, E.C. Slater Institute, Amsterdam, The Netherlands.
EMBO J. 1995 Jul 17;14(14):3472-9. doi: 10.1002/j.1460-2075.1995.tb07353.x.
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.
肉碱乙酰转移酶(CAT)存在于以油酸培养的酿酒酵母的线粒体和过氧化物酶体中。这两种蛋白质由同一个基因YCAT编码,该基因编码的蛋白质在N端具有线粒体靶向信号(MTS),在C端具有1型过氧化物酶体靶向信号(PTS-1)。删除这两个基序后发现存在一个额外的内部靶向序列。通过该内部信号导入CAT被证明依赖于PAS10,PAS10是导入含PTS-1蛋白质所必需的一种蛋白质。利用酵母双杂交技术证明了PAS10与该内部靶向信号之间存在相互作用。在异源启动子后表达YCAT基因导致过氧化物酶体靶向丧失,这表明差异靶向是在转录或翻译水平上受到控制的。对YCAT mRNA 5'端的测定表明,在以油酸培养的细胞中存在在第一个AUG之后起始的YCAT转录本。在以乙酸盐或甘油培养的细胞中几乎不存在这些转录本。我们提出,作为对油酸的响应,会产生较短的转录本,从中翻译出CAT的过氧化物酶体形式,从而产生一种没有MTS的CAT蛋白,该蛋白可靶向过氧化物酶体。