3'UTR AU-Rich Elements (AREs) and the RNA-Binding Protein Tristetraprolin (TTP) Are Not Required for the LPS-Mediated Destabilization of Phospholipase-Cβ-2 mRNA in Murine Macrophages.
3'UTR AU-Rich Elements (AREs) and the RNA-Binding Protein Tristetraprolin (TTP) Are Not Required for the LPS-Mediated Destabilization of Phospholipase-Cβ-2 mRNA in Murine Macrophages.
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DOI：
10.1007/s10753-017-0511-y
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发表时间：
2017-04
期刊：
影响因子：
5.1
通讯作者：
Leibovich SJ
中科院分区：
文献类型：
--
作者：
Shukla S;Elson G;Blackshear PJ;Lutz CS;Leibovich SJ
我们之前已经证明，细菌脂多糖（LPS）介导的磷脂酶- c β-2 （PLCβ-2）表达的抑制参与了腺苷引发的巨噬细胞M1（炎症）向m2样（伤口愈合）表型转换。这种抑制是通过PLCβ-2 mRNA的不稳定介导的。为了研究这种脂多糖介导的不稳定机制，我们研究了rna结合剂（包括microRNAs和rna结合蛋白）在调节编码生长因子、炎症介质和原癌基因的mrna稳定性中的作用。3 ' utr中的富腺苷酸和富尿苷酸（AU）元素（AREs）是rna结合蛋白（包括tristetrprolin （TTP）、HuR和AUF1）以及参与调节mRNA稳定性的microrna的特异性识别位点。在本研究中，我们研究了TTP和AREs在调节PLCβ-2 mRNA稳定性中的作用。将PLCβ-2基因的3'UTR插入pLightswitch荧光素酶报告质粒，转染RAW264.7细胞。LPS抑制荧光素酶的表达。缺乏AREs的突变体3'UTR构建体的荧光素酶表达同样下调，这表明这些区域不是lps介导的PLCβ-2抑制所必需的。在原代小鼠巨噬细胞和RAW264.7细胞中，TTP在LPS的作用下均迅速上调。利用缺乏TTP的小鼠巨噬细胞检测LPS对PLCβ-2的抑制作用。LPS对野生型和TTP−/−巨噬细胞中plc - β-2表达的抑制程度相同。此外，在野生型和TTP - / -巨噬细胞中，lps处理的巨噬细胞中PLCβ-2 mRNA在转录阻断后的衰减率相似，这清楚地表明TTP不参与lps介导的巨噬细胞中PLCβ-2 mRNA的不稳定。
We have shown previously that bacterial lipopolysaccharide (LPS)-mediated suppression of Phospholipase-Cβ-2 (PLCβ-2) expression is involved in M1 (inflammatory) to M2-like (wound healing) phenotypic switching of macrophages triggered by adenosine. This suppression is mediated post-transcriptionally by destabilization of PLCβ-2 mRNA. To investigate the mechanism of this LPS-mediated destabilization, we examined the roles of RNA-binding agents including microRNAs and RNA-binding proteins that are involved in regulating stability of mRNAs encoding growth factors, inflammatory mediators and proto-oncogenes. Adenylate and Uridylate (AU)-rich elements (AREs) in 3′UTRs are specific recognition sites for RNA-binding proteins including Tristetraprolin (TTP), HuR and AUF1, and for microRNAs that are involved in regulating mRNA stability. In this study, we investigated the role of TTP and AREs in regulating PLCβ-2 mRNA stability. The 3′UTR of the PLCβ-2 gene was inserted into the pLightswitch luciferase reporter plasmid and transfected into RAW264.7 cells. LPS suppressed Luciferase expression from this reporter. Luciferase expression from mutant 3′UTR constructs lacking AREs was similarly down-regulated, suggesting that these regions are not required for LPS-mediated suppression of PLCβ-2. TTP was rapidly upregulated in both primary murine macrophages and RAW264.7 cells in response to LPS. Suppression of PLCβ-2 by LPS was examined using macrophages from mice lacking TTP. LPS suppressed PLCβ-2 expression to the same extent in wild type and TTP−/− macrophages. Also, the rate of decay of PLCβ-2 mRNA in LPS-treated macrophages following transcriptional blockade was similar in wild type and TTP−/− macrophages, clearly indicating that TTP is not involved in LPS-mediated destabilization of PLCβ-2 mRNA in macrophages.
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DOI：
10.1016/j.bbagrm.2013.02.003
发表时间：
2013-06
影响因子：
4.7
作者：
Brooks, Seth A.;Blackshear, Perry J.
通讯作者：
Blackshear, Perry J.
影响因子：
11.4
作者：
KEFFER, J;PROBERT, L;KOLLIAS, G
通讯作者：
KOLLIAS, G
影响因子：
3.9
作者：
Carrick, Danielle M.;Blackshear, Perry J.
通讯作者：
Blackshear, Perry J.
影响因子：
6
作者：
Grinberg, Stan;Hasko, Gyorgy;Leibovich, Samuel Joseph
通讯作者：
Leibovich, Samuel Joseph
影响因子：
4.8
作者：
Lai, WS;Carballo, E;Blackshear, PJ
通讯作者：
Blackshear, PJ