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Mouse fat storage-inducing transmembrane protein 2 (FIT2) promotes lipid droplet accumulation in plants

机译:小鼠脂肪储存诱导跨膜蛋白2(FIT2)促进植物中的脂质液滴积聚

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Fat storage-inducing transmembrane protein 2 (FIT2) is an endoplasmic reticulum (ER)-localized protein that plays an important role in lipid droplet (LD) formation in animal cells. However, no obvious homologue of FIT2 is found in plants. Here, we tested the function of FIT2 in plant cells by ectopically expressing mouse (Mus musculus) FIT2 in Nicotiana tabacum suspension-cultured cells, Nicotiana benthamiana leaves and Arabidopsis thaliana plants. Confocal microscopy indicated that the expression of FIT2 dramatically increased the number and size of LDs in leaves of N. benthamiana and Arabidopsis, and lipidomics analysis and mass spectrometry imaging confirmed the accumulation of neutral lipids in leaves. FIT2 also increased seed oil content by similar to 13% in some stable, overexpressing lines of Arabidopsis. When expressed transiently in leaves of N. benthamiana or suspension cells of N. tabacum, FIT2 localized specifically to the ER and was often concentrated at certain regions of the ER that resembled ER-LD junction sites. FIT2 also colocalized at the ER with other proteins known to be involved in triacylglycerol biosynthesis or LD formation in plants, but not with ER resident proteins involved in electron transfer or ER-vesicle exit sites. Collectively, these results demonstrate that mouse FIT2 promotes LD accumulation in plants, a surprising functional conservation in the context of a plant cell given the apparent lack of FIT2 homologues in higher plants. These results suggest also that FIT2 expression represents an effective synthetic biology strategy for elaborating neutral lipid compartments in plant tissues for potential biofuel or bioproduct purposes.
机译:脂肪储存诱导的跨膜蛋白2(FIT2)是内质网(ER) - 循环蛋白质,其在动物细胞中的脂液滴(LD)形成中起重要作用。然而,在植物中没有发现FIT2的明显同源物。在这里,我们通过异常表达烟草悬浮蛋白悬浮细胞,尼古拉··甘羚叶和拟南芥植物来测试FIT2在植物细胞中FIT2在植物细胞中的功能。共聚焦显微镜表明FIT2的表达显着增加了N.Penthamiana和拟南芥叶片中LD的数量和大小,并且脂质体分析和质谱成像证实了叶片中性脂质的积累。 FIT2在一些稳定的过度表达拟南芥中,也会增加种子油含量与13%相似。当在N.Benthamiana的叶片瞬时表达或N. tabacum的悬浮液时,FIT2特异于ER并且通常集中在ER-LD结位点的某些区域。 FIT2也在ER中与已知参与植物的三酰基甘油生物合成或LD形成的其他蛋白质在ER中覆盖,但不是患有电子转移或ER-囊泡出口位点的ER驻留蛋白。总的来说,这些结果表明,小鼠Fit2促进植物中的LD积累,鉴于高等植物中明显缺乏Fit2同源物的植物细胞的背景下令人惊讶的功能守恒。这些结果表明,FIT2表达表示用于在植物组织中阐述中性脂质隔室的有效合成生物学策略,用于潜在的生物燃料或生物制作目的。

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