Kamisugi Yasuko, Mitsuya Shiro, El-Shami Mahmoud, Knight Celia D, Cuming Andrew C, Baker Alison
Centre for Plant Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, UK.
New Phytol. 2016 Jan;209(2):576-89. doi: 10.1111/nph.13739. Epub 2015 Nov 6.
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.
过氧化物酶体生物发生因子11(PEX11)蛋白存在于酵母、哺乳动物和植物中,在过氧化物酶体形态及过氧化物酶体分裂调控中发挥作用。小立碗藓有六种PEX11亚型,可分为两个亚家族,这与单子叶植物和双子叶植物中的情况相似。我们对Phypa_PEX11 - 1基因进行了靶向基因敲除,并比较了野生型和突变株的形态及细胞表型。突变体生长更缓慢,配子体发育受阻。突变体的绿丝体含有大型细胞结构,排斥所有其他细胞器。荧光报告蛋白的表达显示,突变株的过氧化物酶体大幅增大,直径可达10μm。液泡膜标记物的表达证实,增大的结构不是液泡,也不是因pexophagy而被隔离在液泡内的过氧化物酶体。靶向过氧化物酶体膜的Phypa_PEX11可挽救敲除表型,并与过氧化物酶体膜上的Fission1相互作用。小立碗藓中的PEX11在过氧化物酶体分裂中的功能与其他生物体中的PEX11相似,但突变体表型更为极端且受环境影响,这使得小立碗藓成为研究过氧化物酶体增殖和分裂机制的有力系统。