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首页> 外文期刊>AIDS Research and Human Retroviruses >Tax posttranslational modifications and interaction with calreticulin in MT-2 cells and human peripheral blood mononuclear cells of human t cell lymphotropic virus type-i-Associated myelopathy/tropical spastic paraparesis patients
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Tax posttranslational modifications and interaction with calreticulin in MT-2 cells and human peripheral blood mononuclear cells of human t cell lymphotropic virus type-i-Associated myelopathy/tropical spastic paraparesis patients

机译：在人T细胞淋巴病毒I型相关的脊髓病/热带痉挛性轻瘫患者的MT-2细胞和人外周血单核细胞中征税翻译后修饰以及与钙网蛋白相互作用

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The human retrovirus human T cell lymphotropic virus type-I (HTLV-1) is the etiologic agent of HTLV-1-Associated myelopathy/tropical spastic paraparesis (HAM/TSP). Axonal degeneration in HAM/TSP patients occurs without neuron infection, with the secreted viral Tax protein proposed to be involved. We previously found that Tax secreted into the culture medium of MT-2 cells (HTLV-1-infected cell line) produced neurite retraction in neuroblastoma cells differentiated to neuronal type. To assess the relevance of Tax posttranslational modifications on this effect, we addressed the question of whether Tax secreted by MT-2 cells and peripheral blood mononuclear cells (PBMCs) of HTLV-1-infected subjects is modified. The interaction of Tax with calreticulin (CRT) that modulates intracellular Tax localization and secretion has been described. We studied Tax localization and modifications in MT-2 cells and its interaction with CRT. Intracellular Tax in MT-2 cells was assessed by flow cytometry, corresponding mainly to a 71-kDa protein followed by western blot. This protein reported as a chimera with gp21 viral protein-confirmed by mass spectrometry-showed no ubiquitination or SUMOylation. The Tax-CRT interaction was determined by confocal microscopy and coimmunoprecipitation. Extracellular Tax from HAM/TSP PBMCs is ubiquitinated according to western blot, and its interaction with CRT was shown by coimmunoprecipitation. A positive correlation between Tax and CRT secretion was observed in HAM/TSP PBMCs and asymptomatic carriers. For both proteins inhibitors and activators of secretion showed secretion through the endoplasmic reticulum-Golgi complex. Tax, present in PBMC culture medium, produced neurite retraction in differentiated neuroblastoma cells. These results suggest that Tax, whether ubiquitinated or not, is active for neurite retraction.
机译：人类逆转录病毒I型T细胞淋巴病毒（HTLV-1）是HTLV-1-相关性脊髓病/热带痉挛性轻瘫（HAM / TSP）的病因。 HAM / TSP患者的轴突变性在没有神经元感染的情况下发生，分泌的病毒Tax蛋白被认为参与其中。我们以前发现，分泌到MT-2细胞（HTLV-1感染的细胞系）培养基中的Tax分泌在分化为神经元类型的神经母细胞瘤细胞中产生了神经突退缩。为了评估Tax的翻译后修饰对该效果的相关性，我们解决了被HTLV-1感染的受试者的MT-2细胞和外周血单核细胞（PBMC）分泌的Tax是否被修饰的问题。已经描述了Tax与调节细胞内Tax定位和分泌的钙网蛋白（CRT）的相互作用。我们研究了MT-2细胞中的Tax定位和修饰及其与CRT的相互作用。通过流式细胞术评估MT-2细胞的细胞内税，主要对应于71-kDa蛋白，然后进行蛋白质印迹。据质谱图证实，这种蛋白与gp21病毒蛋白为嵌合体，未显示泛素化或SUMOylation。通过共聚焦显微镜和共免疫沉淀法确定了Tax-CRT相互作用。根据蛋白质印迹，来自HAM / TSP PBMC的细胞外税被泛素化，并且其与CRT的相互作用通过共免疫沉淀显示出来。在HAM / TSP PBMC和无症状携带者中观察到Tax和CRT分泌之间呈正相关。对于蛋白质而言，分泌的抑制剂和激活剂均显示出通过内质网-高尔基体的分泌。 PBMC培养基中存在的Tax在分化的神经母细胞瘤细胞中产生神经突退缩。这些结果表明，Tax，无论是否泛素化，对于神经突收缩都是有效的。

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