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首页> 外文期刊>Molecular and Cellular Biology >Transcriptome-Wide RNA Interaction Profiling Reveals Physical and Functional Targets of hnRNP L in Human T Cells
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Transcriptome-Wide RNA Interaction Profiling Reveals Physical and Functional Targets of hnRNP L in Human T Cells

机译：转录组范围的RNA相互作用分析揭示了人类T细胞中hnRNP L的物理和功能靶标

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The RNA processing factor hnRNP L is required for T cell development and function. However, the spectrum of direct targets of hnRNP L activity in T cells has yet to be defined. In this study, we used cross-linking and immunoprecipitation followed by high-throughput sequencing (CLIP-seq) to identify the RNA binding sites of hnRNP L within the transcriptomes of human CD4+ and cultured Jurkat T cells. We find that hnRNP L binds preferentially to transcripts encoding proteins involved in RNA processing and in Wnt and T cell receptor (TCR) signaling. This binding is largely conserved across both quiescent and activated T cells, in agreement with the critical role of hnRNP L throughout T cell biology. Importantly, based on the binding profile of hnRNP L, we validate numerous instances of hnRNP L-dependent alternative splicing of genes critical to T cell function. We further show that alternative exons with weak 5′ splice site sequences specifically show a strong correlation between hnRNP L binding and hnRNP L-dependent splicing regulation. Together, these data provide the first transcriptome-wide analysis of the RNA targets of hnRNP L in lymphoid cells and add to the functional understanding of hnRNP L in human biology.
机译：T细胞发育和功能需要RNA加工因子hnRNPL。但是，尚未确定T细胞中hnRNP L活性的直接靶标范围。在这项研究中，我们使用交联和免疫沉淀法，然后通过高通量测序（CLIP-seq）来鉴定人CD4 + 转录组和培养的Jurkat T细胞内hnRNP L的RNA结合位点。我们发现hnRNP L优先绑定到参与RNA处理以及Wnt和T细胞受体（TCR）信号传导的编码蛋白质的转录本。与hnRNP L在整个T细胞生物学中的关键作用相一致，这种结合在静止T细胞和活化T细胞之间都非常保守。重要的是，基于hnRNP L的结合情况，我们验证了对T细胞功能至关重要的基因hnRNP L依赖的选择性剪接的许多实例。我们进一步表明，具有弱5'剪接位点序列的替代外显子特别显示hnRNP L结合与hnRNP L依赖的剪接调控之间的强相关性。总之，这些数据提供了对淋巴样细胞中hnRNP L RNA靶标的首次转录组范围分析，并增加了对hnRNP L在人类生物学中的功能性了解。

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