Quantification of transcriptome changes to investigate the role of glucocorticoid receptor-RNA binding during dexamethasone treatment.
Quantification of transcriptome changes to investigate the role of glucocorticoid receptor-RNA binding during dexamethasone treatment.
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DOI：
10.1186/s13104-023-06446-4
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发表时间：
2023-08-22
影响因子：
1.8
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中科院分区：
文献类型：
--
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糖皮质激素受体（GR）是一种被广泛研究的配体激活转录因子，也是抗炎治疗的常见靶点。最近，一些研究引起了人们对GR与RNA而非DNA结合的影响以及这种活性对GR功能的潜在影响的关注。我们研究的目的是通过测量在表现出RNA亲和力降低的GR突变体存在时糖皮质激素驱动的转录组的变化，进一步表征GR功能与RNA结合之间的关系。在含有GR构建体（GR- halotag）的三个细胞系中，GR被激活。其中一个细胞系含有野生型GR-HaloTag。另一种含有GR-HaloTag，其突变降低了RNA亲和力，并略微降低了DNA亲和力。第三个细胞系含有GR-HaloTag，其突变仅略微降低了DNA亲和力。所有三种细胞系均用地塞米松（一种GR激动剂）处理。每隔一小时采集RNA-seq样本，持续3小时。此外，转录组定量通过使用4-硫脲标记地塞米松治疗最后一小时转录的rna来完成。然后对这些标记的rna进行纯化和测序。该数据集是GR的第一个同类数据集，包含了对GR RNA结合功能的有价值的见解。
The glucocorticoid receptor (GR) is a well-studied, ligand-activated transcription factor and a common target of anti-inflammatory treatments. Recently, several studies have drawn attention the effects of binding of GR to RNA rather than DNA and the potential implications of this activity for GR function. The objective of our study was to further characterize the relationship between GR function and RNA binding by measuring changes in the glucocorticoid-driven transcriptome in the presence of a GR mutant that exhibited reduced RNA affinity. GR was activated in three cell lines containing GR constructs (GR-HaloTag). One of the cell lines contained a wild-type GR-HaloTag. Another contained GR-HaloTag with a mutation that reduced RNA affinity and slightly reduced DNA affinity. The third cell line contained GR-HaloTag with a mutation that only slightly reduced DNA affinity. All three cell lines were treated with dexamethasone, a GR agonist. RNA-seq samples were collected every hour for 3 h. Moreover, transcriptome quantification was accomplished via labeling of RNAs transcribed in the final hour of dexamethasone treatment using 4-thiouridine. These labeled RNAs were then purified and sequenced. This data set is the first of its kind for GR and contains valuable insights into the function of RNA binding by GR.
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通讯作者：
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