Marzioch M, Erdmann R, Veenhuis M, Kunau W H
Institut für Physiologische Chemie, Ruhr-Universität Bochum, Germany.
EMBO J. 1994 Oct 17;13(20):4908-18. doi: 10.1002/j.1460-2075.1994.tb06818.x.
To identify components of the peroxisomal import pathway in yeast, we have isolated pas mutants affected in peroxisome biogenesis. Two mutants assigned to complementation group 7 define a new gene, PAS7, whose product is necessary for import of thiolase, a PTS2-containing protein, but not for that of SKL (PTS1)-containing proteins, into peroxisomes. We have cloned PAS7 by complementation of the oleic acid non-utilizing phenotype of the pas7-1 strain. The DNA sequence predicts a 42.3 kDa polypeptide of 375 amino acids encoding a novel member of the beta-transducin related (WD-40) protein family. A Myc epitope-tagged Pas7p, expressed under the control of the CUP1 promotor, was functionally active. Subcellular localization studies revealed that in the presence of thiolase this epitope-tagged Pas7p in part associates with peroxisomes. However, in a thiolase-deficient mutant, Pas7p was entirely found in the cytoplasm. We suggest that Pas7p mediates the binding of thiolase to these organelles.
为了鉴定酵母中过氧化物酶体输入途径的组成成分,我们分离了在过氧化物酶体生物发生过程中受到影响的pas突变体。被归入互补群7的两个突变体定义了一个新基因PAS7,其产物对于硫解酶(一种含PTS2的蛋白质)进入过氧化物酶体是必需的,但对于含SKL(PTS1)的蛋白质进入过氧化物酶体则不是必需的。我们通过互补pas7-1菌株不利用油酸的表型克隆了PAS7。DNA序列预测该基因编码一个由375个氨基酸组成、分子量为42.3 kDa的多肽,它是β-转导蛋白相关(WD-40)蛋白家族的一个新成员。在CUP1启动子控制下表达的带有Myc表位标签的Pas7p具有功能活性。亚细胞定位研究表明,在有硫解酶存在的情况下,这种带有表位标签的Pas7p部分地与过氧化物酶体相关联。然而,在硫解酶缺陷型突变体中,Pas7p完全存在于细胞质中。我们认为Pas7p介导硫解酶与这些细胞器的结合。