Kollipara Laxmikanth, Buchkremer Stephan, Coraspe José Andrés González, Hathazi Denisa, Senderek Jan, Weis Joachim, Zahedi René P, Roos Andreas
Leibniz-Institut für Analytische Wissenschaften-ISAS -e.V., 44227 Dortmund, Germany.
Institute of Neuropathology, University Hospital Aachen, RWTH Aachen, 5274 Aachen, Germany.
Oncotarget. 2017 Jul 28;8(40):68493-68516. doi: 10.18632/oncotarget.19663. eCollection 2017 Sep 15.
SIL1 is a ubiquitous protein of the Endoplasmic Reticulum (ER) acting as a co-chaperone for the ER-resident chaperone, BiP. Recessive mutations of the corresponding gene lead to vulnerability of skeletal muscle and central nervous system in man (Marinesco-Sjögren syndrome; MSS) and mouse. However, it is still unclear how loss of ubiquitous SIL1 leads to selective vulnerability of the nervous system and skeletal muscle whereas other cells and organs are protected from clinical manifestations. In this study we aimed to disentangle proteins participating in selective vulnerability of SIL1-deficient cells and tissues: morphological examination of MSS patient-derived lymphoblastoid cells revealed altered organelle structures (ER, nucleus and mitochondria) thus showing subclinical vulnerability. To correlate structural perturbations with biochemical changes and to identify proteins potentially preventing phenotypical manifestation, proteomic studies have been carried out. Results of proteomic profiling are in line with the morphological findings and show affection of nuclear, mitochondrial and cytoskeletal proteins as well as of such responsible for cellular viability. Moreover, expression patterns of proteins known to be involved in neuromuscular disorders or in development and function of the nervous system were altered. Paradigmatic findings were confirmed by immunohistochemistry of splenic lymphocytes and the cerebellum of SIL1-deficient mice. Ataxin-10, identified with increased abundance in our proteome profile, is necessary for the neuronal survival but also controls muscle fiber apoptosis, thus declaring this protein as a plausible candidate for selective tissue vulnerability. Our combined results provide first insights into the molecular causes of selective cell and tissue vulnerability defining the MSS phenotype.
SIL1是一种内质网(ER)中普遍存在的蛋白质,作为内质网驻留伴侣蛋白BiP的共伴侣蛋白发挥作用。相应基因的隐性突变会导致人类( Marinesco-Sjögren综合征;MSS)和小鼠的骨骼肌和中枢神经系统易损。然而,目前仍不清楚普遍存在的SIL1缺失如何导致神经系统和骨骼肌的选择性易损,而其他细胞和器官却没有出现临床表现。在本研究中,我们旨在厘清参与SIL1缺陷细胞和组织选择性易损的蛋白质:对MSS患者来源的淋巴母细胞进行形态学检查,发现细胞器结构(内质网、细胞核和线粒体)发生改变,从而显示出亚临床易损性。为了将结构扰动与生化变化相关联,并鉴定可能预防表型表现的蛋白质,我们进行了蛋白质组学研究。蛋白质组学分析结果与形态学发现一致,显示核蛋白、线粒体蛋白和细胞骨架蛋白以及负责细胞活力的蛋白受到影响。此外,已知参与神经肌肉疾病或神经系统发育和功能的蛋白质的表达模式也发生了改变。通过对SIL1缺陷小鼠的脾淋巴细胞和小脑进行免疫组织化学分析,证实了典型的研究结果。在我们的蛋白质组图谱中丰度增加的Ataxin-10对神经元存活是必需的,但也控制肌纤维凋亡,因此表明该蛋白是选择性组织易损的一个合理候选蛋白。我们的综合研究结果首次揭示了定义MSS表型的选择性细胞和组织易损的分子原因。