Science for Life Laboratory, Royal Institute of Technology, Stockholm, Sweden.
Mol Syst Biol. 2010 Dec 21;6:450. doi: 10.1038/msb.2010.106.
An essential question in human biology is how cells and tissues differ in gene and protein expression and how these differences delineate specific biological function. Here, we have performed a global analysis of both mRNA and protein levels based on sequence-based transcriptome analysis (RNA-seq), SILAC-based mass spectrometry analysis and antibody-based confocal microscopy. The study was performed in three functionally different human cell lines and based on the global analysis, we estimated the fractions of mRNA and protein that are cell specific or expressed at similar/different levels in the cell lines. A highly ubiquitous RNA expression was found with >60% of the gene products detected in all cells. The changes of mRNA and protein levels in the cell lines using SILAC and RNA ratios show high correlations, even though the genome-wide dynamic range is substantially higher for the proteins as compared with the transcripts. Large general differences in abundance for proteins from various functional classes are observed and, in general, the cell-type specific proteins are low abundant and highly enriched for cell-surface proteins. Thus, this study shows a path to characterize the transcriptome and proteome in human cells from different origins.
一个在人类生物学中的重要问题是细胞和组织在基因和蛋白质表达上的差异,以及这些差异如何界定特定的生物学功能。在这里,我们基于基于序列的转录组分析(RNA-seq)、SILAC 基于质谱分析和基于抗体的共聚焦显微镜,对 mRNA 和蛋白质水平进行了全面分析。该研究在三种功能不同的人类细胞系中进行,基于全面分析,我们估计了在细胞系中细胞特异性表达或表达水平相似/不同的 mRNA 和蛋白质的分数。我们发现高度普遍的 RNA 表达,超过 60%的基因产物在所有细胞中都有检测到。使用 SILAC 和 RNA 比率的细胞系中 mRNA 和蛋白质水平的变化显示出高度相关性,尽管与转录物相比,蛋白质的全基因组动态范围要高得多。从各种功能类别观察到蛋白质丰度的巨大总体差异,并且通常情况下,细胞类型特异性蛋白质的丰度较低,富含细胞表面蛋白。因此,这项研究为从不同来源的人类细胞中表征转录组和蛋白质组提供了一条途径。