Saberianfar Reza, Sattarzadeh Amirali, Joensuu Jussi J, Kohalmi Susanne E, Menassa Rima
Agriculture and Agri-Food CanadaLondon, ON, Canada; Department of Biology, University of Western OntarioLondon, ON, Canada.
Department of Molecular Biology and Genetics, Cornell University Ithaca, NY, USA.
Front Plant Sci. 2016 May 23;7:693. doi: 10.3389/fpls.2016.00693. eCollection 2016.
Protein bodies (PBs) are organelles found in seeds whose main function is the storage of proteins that are used during germination for sustaining growth. PBs can also be induced to form in leaves when foreign proteins are produced at high levels in the endoplasmic reticulum (ER) and when fused to one of three tags: Zera®, elastin-like polypeptides (ELP), or hydrophobin-I (HFBI). In this study, we investigate the differences between ELP, HFBI and Zera PB formation, packing, and communication. Our results confirm the ER origin of all three fusion-tag-induced PBs. We show that secretory pathway proteins can be sequestered into all types of PBs but with different patterns, and that different fusion tags can target a specific protein to different PBs. Zera PBs are mobile and dependent on actomyosin motility similar to ELP and HFBI PBs. We show in vivo trafficking of proteins between PBs using GFP photoconversion. We also show that protein trafficking between ELP or HFBI PBs is faster and proteins travel further when compared to Zera PBs. Our results indicate that fusion-tag-induced PBs do not represent terminally stored cytosolic organelles, but that they form in, and remain part of the ER, and dynamically communicate with each other via the ER. We hypothesize that the previously documented PB mobility along the actin cytoskeleton is associated with ER movement rather than independent streaming of detached organelles.
蛋白体(PBs)是种子中发现的细胞器,其主要功能是储存蛋白质,这些蛋白质在种子萌发时用于维持生长。当外源蛋白在内质网(ER)中高水平产生并与三种标签之一(Zera®、弹性蛋白样多肽(ELP)或疏水蛋白-I(HFBI))融合时,也可诱导叶片中形成蛋白体。在本研究中,我们调查了ELP、HFBI和Zera蛋白体形成、堆积和通讯之间的差异。我们的结果证实了所有三种融合标签诱导的蛋白体均起源于内质网。我们表明,分泌途径蛋白可以被隔离到所有类型的蛋白体中,但模式不同,并且不同的融合标签可以将特定蛋白靶向到不同的蛋白体。Zera蛋白体是可移动的,并且与ELP和HFBI蛋白体一样依赖于肌动球蛋白的运动性。我们使用绿色荧光蛋白(GFP)光转化在体内展示了蛋白体之间的蛋白运输。我们还表明,与Zera蛋白体相比,ELP或HFBI蛋白体之间的蛋白运输更快,蛋白移动得更远。我们的结果表明,融合标签诱导的蛋白体并不代表终末储存的胞质细胞器,而是在内质网中形成并仍然是内质网的一部分,并通过内质网彼此动态通讯。我们推测,先前记录的蛋白体沿肌动蛋白细胞骨架的移动性与内质网的移动有关,而不是独立细胞器的流动。