Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
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DOI：
10.1093/nar/gkf682
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发表时间：
2002-12-15
影响因子：
14.9
通讯作者：
Bohjanen, PR
中科院分区：
文献类型：
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作者：
Raghavan, A;Ogilvie, RL;Bohjanen, PR
为了更好地了解mRNA衰减在调节基因表达中的作用，我们使用微阵列技术测量了静息和活化T淋巴细胞的mRNA衰减率。纯化的人T淋巴细胞分别用培养基、抗cd3抗体或抗cd3和抗cd28抗体联合刺激3小时。加入放线菌素D来阻止转录，并在2小时内的离散时间点收集细胞总RNA。使用Affymetrix寡核苷酸阵列分析来自每个点的RNA，并使用一阶衰变模型来确定大约6000个表达转录本的半衰期。我们发现了数百种编码重要调节蛋白的短寿命转录本，包括细胞因子、细胞表面受体、信号转导调节因子、转录因子、细胞周期调节因子和凋亡调节因子。这些短寿命转录本中大约有100个含有类似于ares的序列。我们还发现了许多转录本在mRNA衰变中表现出刺激依赖性的变化。特别是，我们确定了数百个转录本，其稳态水平在T细胞激活后被抑制，在静息状态下不稳定或在细胞激活后不稳定。因此，mRNA的快速降解似乎是以激活依赖的方式关闭基因表达的重要机制。
We used microarray technology to measure mRNA decay rates in resting and activated T lymphocytes in order to better understand the role of mRNA decay in regulating gene expression. Purified human T lymphocytes were stimulated for 3 h with medium alone, with an anti-CD3 antibody, or with a combination of anti-CD3 and anti-CD28 antibodies. Actinomycin D was added to arrest transcription, and total cellular RNA was collected at discrete time points over a 2 h period. RNA from each point was analyzed using Affymetrix oligonucleotide arrays and a first order decay model was used to determine the half-lives of approximately 6000 expressed transcripts. We identified hundreds of short-lived transcripts encoding important regulatory proteins including cytokines, cell surface receptors, signal transduction regulators, transcription factors, cell cycle regulators and regulators of apoptosis. Approximately 100 of these short-lived transcripts contained ARE-like sequences. We also identified numerous transcripts that exhibited stimulus-dependent changes in mRNA decay. In particular, we identified hundreds of transcripts whose steady-state levels were repressed following T cell activation and were either unstable in the resting state or destabilized following cellular activation. Thus, rapid mRNA degradation appears to be an important mechanism for turning gene expression off in an activation-dependent manner.