Plant Cell Biology, Biological and Medical Sciences, Oxford Brookes University, Oxford, OX3 0BP, UK.
School of Life Sciences, University of Warwick, Gibbet Hill, Coventry, CV4 7AL, UK.
New Phytol. 2018 Aug;219(3):990-1004. doi: 10.1111/nph.15228. Epub 2018 May 25.
The plant endoplasmic reticulum (ER) is crucial to the maintenance of cellular homeostasis. The ER consists of a dynamic and continuously remodelling network of tubules and cisternae. Several conserved membrane proteins have been implicated in formation and maintenance of the ER network in plants, such as RHD3 and the reticulon proteins. Despite the recent work in mammalian and yeast cells, the detailed molecular mechanisms of ER network organization in plants remain largely unknown. Recently, novel ER network-shaping proteins called Lunapark (LNP) have been identified in yeast and mammalian cells. Here we identify two Arabidopsis LNP homologues and investigate their subcellular localization via confocal microscopy and potential function in shaping the ER network using protein-protein interaction assays and mutant analysis. We show that AtLNP1 overexpression in tobacco leaf epidermal cells mainly labels cisternae in the ER network, whereas AtLNP2 labels the whole ER. Overexpression of LNP proteins results in an increased abundance of ER cisternae and lnp1 and lnp1lnp2 amiRNA lines display a reduction in cisternae and larger polygonal areas. Thus, we hypothesize that AtLNP1 and AtLNP2 are involved in determining the network morphology of the plant ER, possibly by regulating the formation of ER cisternae.
植物内质网(ER)对维持细胞内稳态至关重要。ER 由一个动态的、不断重塑的小管和小泡网络组成。几种保守的膜蛋白已被牵连到植物 ER 网络的形成和维持中,如 RHD3 和 reticulon 蛋白。尽管最近在哺乳动物和酵母细胞中进行了研究,但植物 ER 网络组织的详细分子机制在很大程度上仍不清楚。最近,在酵母和哺乳动物细胞中发现了一种称为 Lunapark(LNP)的新型 ER 网络成型蛋白。在这里,我们鉴定了两个拟南芥 LNP 同源物,并通过共聚焦显微镜研究了它们的亚细胞定位,以及通过蛋白-蛋白相互作用分析和突变分析研究了它们在塑造 ER 网络中的潜在功能。我们发现 AtLNP1 在烟草叶表皮细胞中的过表达主要标记 ER 网络中的小泡,而 AtLNP2 标记整个 ER。LNP 蛋白的过表达导致 ER 小泡的丰度增加,而 lnp1 和 lnp1lnp2 amiRNA 系显示小泡减少和更大的多边形区域。因此,我们假设 AtLNP1 和 AtLNP2 参与决定植物 ER 的网络形态,可能通过调节 ER 小泡的形成。