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Giant peroxisomes in a moss (Physcomitrella patens) peroxisomal biogenesis factor 11 mutant

机译:巨型过氧化物酶体在苔藓(physcomitrella patens)过氧化物酶体生物合成因子11突变体

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摘要

Peroxisomal biogenesis factor 11 (PEX11) proteins are found in yeasts, mammals and plants, and play a role in peroxisome morphology and regulation of peroxisome division. The moss Physcomitrella patens has six PEX11 isoforms which fall into two subfamilies, similar to those found in monocots and dicots. We carried out targeted gene disruption of the Phypa_PEX11-1 gene and compared the morphological and cellular phenotypes of the wild-type and mutant strains. The mutant grew more slowly and the development of gametophores was retarded. Mutant chloronemal filaments contained large cellular structures which excluded all other cellular organelles. Expression of fluorescent reporter proteins revealed that the mutant strain had greatly enlarged peroxisomes up to 10 μm in diameter. Expression of a vacuolar membrane marker confirmed that the enlarged structures were not vacuoles, or peroxisomes sequestered within vacuoles as a result of pexophagy. Phypa_PEX11 targeted to peroxisome membranes could rescue the knock out phenotype and interacted with Fission1 on the peroxisome membrane. Moss PEX11 functions in peroxisome division similar to PEX11 in other organisms but the mutant phenotype is more extreme and environmentally determined, making P. patens a powerful system in which to address mechanisms of peroxisome proliferation and division.
机译:过氧化物酶体生物生成因子11(PEX11)蛋白存在于酵母,哺乳动物和植物中,并在过氧化物酶体形态和过氧化物酶体分裂的调控中发挥作用。苔藓假单胞菌有六个PEX11亚型,分为两个亚科,类似于在单子叶植物和双子叶植物中发现的亚型。我们对Phypa_PEX11-1基因进行了有针对性的基因破坏,并比较了野生型和突变菌株的形态和细胞表型。突变体生长较慢,配子体的发育受阻。突变的氯nemal细丝包含大的细胞结构,排除了所有其他细胞器。荧光报告蛋白的表达表明该突变菌株的过氧化物酶体极大地扩大了直径,最大可达10μm。液泡膜标记物的表达证实了扩大结构不是液泡,或由于过氧化物酶病而使过氧化物酶体隔离在液泡内。靶向过氧化物酶体膜的Phypa_PEX11可以挽救基因敲除表型,并与过氧化物酶体膜上的Fission1相互作用。苔藓PEX11在过氧化物酶体分裂中的功能与其他生物体中的PEX11类似,但突变表型更为极端且由环境决定,这使P. patens成为解决过氧化物酶体增殖和分裂机制的强大系统。

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