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Peroxisome biogenesis occurs in an unsynchronized manner in close association with the endoplasmic reticulum in temperature-sensitive Yarrowia lipolytica Pex3p mutants

机译:过氧化物酶体生物发生以不同步的方式与温度敏感的解脂耶氏酵母Pex3p突变体的内质网密切相关

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Pex3p is a peroxisomal integral membrane protein required early in peroxisome biogenesis, and Pex3p-deficient cells lack identifiable peroxisomes. Two temperature-sensitive pex3 mutant strains of the yeast Yarrowia lipolytica were made to investigate the role of Pex3p in the early stages of peroxisome biogenesis. In glucose medium at 16degreesC, these mutants underwent de novo peroxisome biogenesis and exhibited early matrix protein sequestration into peroxisome-like structures found at the endoplasmic reticulum-rich periphery of cells or sometimes associated with nuclei. The de novo peroxisome biogenesis seemed unsynchronized, with peroxisomes occurring at different stages of development both within cells and between cells. Cells with peripheral nascent peroxisomes and cells with structures morphologically distinct from peroxisomes, such as semi/circular tubular structures that immunostained with antibodies to peroxisomal matrix proteins and to the endoplasmic reticulum-resident protein Kar2p, and that surrounded lipid droplets, were observed during up-regulation of peroxisome biogenesis in cells incubated in oleic acid medium at 16degreesC. These structures were not detected in wild-type or Pex3p-deficient cells. Their role in peroxisome biogenesis remains unclear. Targeting of peroxisomal matrix proteins to these structures suggests that Pex3p directly or indirectly sequesters components of the peroxisome biogenesis machinery. Such a role is consistent with Pex3p overexpression producing cells with fewer, larger, and clustered peroxisomes. [References: 46]
机译:Pex3p是过氧化物酶体生物发生早期所需的过氧化物酶体整合膜蛋白,而缺乏Pex3p的细胞缺乏可识别的过氧化物酶体。制备了两个解脂耶氏酵母酵母的温度敏感型pex3突变菌株,以研究Pex3p在过氧化物酶体生物合成的早期阶段中的作用。在16°C的葡萄糖培养基中,这些突变体从头进行过氧化物酶体生物合成,并表现出早期的基质蛋白螯合成过氧化物酶体样结构,存在于细胞内质网富集的外围或有时与细胞核相关。从头过氧化物酶体的生物发生似乎是不同步的,过氧化物酶体发生在细胞内和细胞之间的不同发育阶段。在上升过程中,观察到具有周边新生过氧化物酶体的细胞和形态与过氧化物酶体不同的细胞,如半/圆形管状结构,其被过氧化物酶体基质蛋白和内质网驻留蛋白Kar2p抗体免疫染色,并围绕脂质滴。于16°C油酸培养基中孵育的细胞中过氧化物酶体生物合成的调控。在野生型或Pex3p缺陷型细胞中未检测到这些结构。它们在过氧化物酶体生物发生中的作用尚不清楚。过氧化物酶体基质蛋白靶向这些结构表明Pex3p直接或间接隔离过氧化物酶体生物发生机制的组件。这种作用与Pex3p过表达产生的细胞具有更少,更大和成簇的过氧化物酶体一致。 [参考:46]

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