O-glycan initiation directs distinct biological pathways and controls epithelial differentiation
O-glycan initiation directs distinct biological pathways and controls epithelial differentiation
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DOI：
10.15252/embr.201948885
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发表时间：
2020-04-23
期刊：
影响因子：
7.7
通讯作者：
Wandall, Hans H.
中科院分区：
文献类型：
--
作者：
Bagdonaite, Ieva;Pallesen, Emil M. H.;Wandall, Hans H.
翻译后修饰（PTMs）极大地扩展了蛋白质活性调节的功能和潜力，O-糖基化是最丰富和多样的PTMs之一。O-GalNAc糖基化的起始受20种不同的GalNAc-转移酶（GalNAc-Ts）调节，并且个体GalNAc-Ts中的缺陷与人类疾病相关，在模式生物中引起微妙但不同的表型。在这里，我们产生了一组等基因角质形成细胞系缺乏三个占主导地位的和差异表达的GalNAc-T。通过角质形成细胞形成上皮的能力，我们研究了单个GalNAc-Ts损失的表型后果。此外，我们通过全球转录组学，差异糖蛋白质组学和差异磷酸蛋白质组学分析探测细胞反应。我们证明，个别GalNAc-T亚型的损失导致不同的上皮表型，通过其对特定的生物学途径的影响; GalNAc-T1目标与内膜系统的组件，GalNAc-T2目标与细胞-ECM粘附，和GalNAc-T3目标与上皮分化。因此，GalNAc-T同种型在人上皮组织形成期间发挥特定作用。
Post-translational modifications (PTMs) greatly expand the function and potential for regulation of protein activity, and O-glycosylation is among the most abundant and diverse PTMs. Initiation of O-GalNAc glycosylation is regulated by 20 distinct GalNAc-transferases (GalNAc-Ts), and deficiencies in individual GalNAc-Ts are associated with human disease, causing subtle but distinct phenotypes in model organisms. Here, we generate a set of isogenic keratinocyte cell lines lacking either of the three dominant and differentially expressed GalNAc-Ts. Through the ability of keratinocytes to form epithelia, we investigate the phenotypic consequences of the loss of individual GalNAc-Ts. Moreover, we probe the cellular responses through global transcriptomic, differential glycoproteomic, and differential phosphoproteomic analyses. We demonstrate that loss of individual GalNAc-T isoforms causes distinct epithelial phenotypes through their effect on specific biological pathways; GalNAc-T1 targets are associated with components of the endomembrane system, GalNAc-T2 targets with cell-ECM adhesion, and GalNAc-T3 targets with epithelial differentiation. Thus, GalNAc-T isoforms serve specific roles during human epithelial tissue formation.