Quantitative proteomic analysis of Myc oncoprotein function
Quantitative proteomic analysis of Myc oncoprotein function
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DOI：
10.1093/emboj/cdf525
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发表时间：
2002-10-01
期刊：
影响因子：
11.4
通讯作者：
Eisenman, RN
中科院分区：
文献类型：
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作者：
Shiio, Y;Donohoe, S;Eisenman, RN
本研究应用一种新的定量蛋白质组学技术来分析哺乳动物细胞中Myc癌蛋白的功能。采用同位素编码亲和标签（ICAT™）试剂标记和串联质谱法，将大鼠myc-null细胞中蛋白质表达的整体模式与myc-plus细胞（已引入myc的myc-null细胞）进行比较，以生成差异蛋白质表达目录。鉴定出许多功能相关蛋白质之间的表达差异，包括 myc-plus 细胞中蛋白酶的减少、蛋白质合成途径的诱导和合成代谢酶的上调，预计这些差异将导致细胞质量增加（细胞生长）。此外，在 myc-plus 细胞中观察到粘附分子、肌动蛋白网络蛋白和 Rho 通路蛋白水平降低，导致粘着斑和肌动蛋白应力纤维减少以及形态改变。这些效应取决于高度保守的 Myc Box II 区域。我们的结果揭示了 Myc 的一种新的细胞骨架功能，并表明在哺乳动物细胞中进行定量全蛋白质组分析的可行性。
This study applies a new quantitative proteomics technology to the analysis of the function of the Myc oncoprotein in mammalian cells. Employing isotope-coded affinity tag (ICAT(TM)) reagent labeling and tandem mass spectrometry, the global pattern of protein expression in rat myc-null cells was compared with that of myc-plus cells (myc-null cells in which myc has been introduced) to generate a differential protein expression catalog. Expression differences among many functionally related proteins were identified, including reduction of proteases, induction of protein synthesis pathways and upregulation of anabolic enzymes in myc-plus cells, which are predicted to lead to increased cell mass (cell growth). In addition, reduction in the levels of adhesion molecules, actin network proteins and Rho pathway proteins were observed in myc-plus cells, leading to reduced focal adhesions and actin stress fibers as well as altered morphology. These effects are dependent on the highly conserved Myc Box II region. Our results reveal a novel cytoskeletal function for Myc and indicate the feasibility of quantitative whole-proteome analysis in mammalian cells.