Lay Dorothee, L Grosshans Bianka, Heid Hans, Gorgas Karin, Just Wilhelm W
Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, D-69120 Heidelberg, Germany.
J Biol Chem. 2005 Oct 14;280(41):34489-99. doi: 10.1074/jbc.M503497200. Epub 2005 Aug 12.
We have analyzed in vitro the binding characteristics of members of the ADP-ribosylation factor (ARF) family of proteins to a highly purified rat liver peroxisome preparation void of Golgi membranes and studied in vivo a role these proteins play in the proliferation of yeast peroxisomes. Although both ARF1 and ARF6 were found on peroxisomes, coatomer recruitment only depended on ARF1-GTP. Recruitment of ARF1 and coatomer to peroxisomes was significantly affected both by pretreating the animals with peroxisome proliferators and by ATP and a cytosolic fraction designated the intermediate pool fraction depleted of ARF and coatomer. In the presence of ATP, the concentrations of ARF1 and coatomer on peroxisomes were reduced, whereas intermediate pool fraction led to a concentration-dependent decrease in ARF and increase in coatomer. Brefeldin A, a fungal toxin that is known to reduce ARF1 binding to Golgi membranes, did not affect ARF1 binding to peroxisomes. In Saccharomyces cerevisiae, both ScARF1 and ScARF3, the yeast orthologs of mammalian ARF1 and ARF6, were implicated in the control of peroxisome proliferation. ScARF1 regulated this process in a positive manner, and ScARF3 regulated it in a negative manner.
我们在体外分析了ADP核糖基化因子（ARF）蛋白家族成员与不含高尔基体膜的高度纯化大鼠肝脏过氧化物酶体制剂的结合特性，并在体内研究了这些蛋白在酵母过氧化物酶体增殖中所起的作用。虽然在过氧化物酶体上发现了ARF1和ARF6，但衣被蛋白的募集仅依赖于ARF1-GTP。用过氧化物酶体增殖剂预处理动物以及ATP和一种称为中间池组分的不含ARF和衣被蛋白的胞质组分，均显著影响ARF1和衣被蛋白向过氧化物酶体的募集。在ATP存在的情况下，过氧化物酶体上ARF1和衣被蛋白的浓度降低，而中间池组分导致ARF浓度依赖性降低和衣被蛋白增加。布雷菲德菌素A是一种已知可减少ARF1与高尔基体膜结合的真菌毒素，它不影响ARF1与过氧化物酶体的结合。在酿酒酵母中，哺乳动物ARF1和ARF6的酵母同源物ScARF1和ScARF3均参与过氧化物酶体增殖的调控。ScARF1以正向方式调节这一过程，而ScARF3以负向方式调节。
