Co-regulation of intragenic microRNA miR-153 and its host gene Ia-2 β: identification of miR-153 target genes with functions related to IA-2β in pancreas and brain.
Co-regulation of intragenic microRNA miR-153 and its host gene Ia-2 β: identification of miR-153 target genes with functions related to IA-2β in pancreas and brain.
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DOI：
10.1007/s00125-013-2901-5
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发表时间：
2013-07
期刊：
影响因子：
8.2
通讯作者：
De Strooper, B.
中科院分区：
文献类型：
--
作者：
Mandemakers, W.;Abuhatzira, L.;Xu, H.;Caromile, L. A.;Hebert, S. S.;Snellinx, A.;Morais, V. A.;Matta, S.;Cai, T.;Notkins, A. L.;De Strooper, B.
我们分析了miR-153的基因组组织，miR-153是嵌入在编码两种主要的1型糖尿病自身抗原，胰岛相关蛋白(IA)-2和IA-2β的基因中的微小rna。我们还发现了与IA-2β定位和功能相关的miR-153靶基因。采用生物信息学方法鉴定miR-153的基因组组织。为了分析miR-153和Ia-2β的共同调控，在葡萄糖刺激实验后，在MIN6B细胞和分离的小鼠胰岛，以及野生型Ia-2（也称为Ptprn）， Ia-2β单敲除和Ia-2/Ia-2β双敲除小鼠脑和胰岛中进行了miR-153和Ia-2β（也称为Ptprn）的定量PCR分析。对miR-153靶基因进行生物信息学鉴定，并通过荧光素酶报告基因检测、western blotting和定量PCR进行验证。miR-153的两个拷贝，miR-153-1和miR-153-2，分别定位于Ia-2和Ia-2β的内含子19中。在啮齿类动物中，只有miR-153-2是保守的。我们证明了miR-153-2和Ia-2β在啮齿类动物中的表达是部分共调节的，正如在Ia-2β敲除和Ia-2/Ia-2β双敲除小鼠中miR-153表达水平的显著降低所证明的那样。miR-153水平在Ia-2敲除小鼠中不受影响。此外，葡萄糖刺激增加了Ia-2和Ia-2β的表达，也显著增加了miR-153的表达。体外葡萄糖刺激后，miR-153的几个预测靶标降低，这与miR-153水平的升高相关。这项研究表明miR-153、IA-2β和miR-153靶基因参与了一个调控网络，这可能与胰岛素和神经递质释放有关。本文的在线版本（doi:10.1007/s00125-013-2901-5）包含同行评审但未经编辑的补充材料，授权用户可使用。
We analysed the genomic organisation of miR-153, a microRNA embedded in genes that encode two of the major type 1 diabetes autoantigens, islet-associated protein (IA)-2 and IA-2β. We also identified miR-153 target genes that correlated with IA-2β localisation and function. A bioinformatics approach was used to identify miR-153’s genomic organisation. To analyse the co-regulation of miR-153 and IA-2β, quantitative PCR analysis of miR-153 and Ia-2β (also known as Ptprn2) was performed after a glucose stimulation assay in MIN6B cells and isolated murine pancreatic islets, and also in wild-type Ia-2 (also known as Ptprn), Ia-2β single knockout and Ia-2/Ia-2β double knockout mouse brain and pancreatic islets. Bioinformatics identification of miR-153 target genes and validation via luciferase reporter assays, western blotting and quantitative PCR were also carried out. Two copies of miR-153, miR-153-1 and miR-153-2, are localised in intron 19 of Ia-2 and Ia-2β, respectively. In rodents, only miR-153-2 is conserved. We demonstrated that expression of miR-153-2 and Ia-2β in rodents is partially co-regulated as demonstrated by a strong reduction of miR-153 expression levels in Ia-2β knockout and Ia-2/Ia-2β double knockout mice. miR-153 levels were unaffected in Ia-2 knockout mice. In addition, glucose stimulation, which increases Ia-2 and Ia-2β expression, also significantly increased expression of miR-153. Several predicted targets of miR-153 were reduced after glucose stimulation in vitro, correlating with the increase in miR-153 levels. This study suggests the involvement of miR-153, IA-2β and miR-153 target genes in a regulatory network, which is potentially relevant to insulin and neurotransmitter release. The online version of this article (doi:10.1007/s00125-013-2901-5) contains peer-reviewed but unedited supplementary material, which is available to authorised users.
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DOI：
10.1152/ajpendo.00262.2010
发表时间：
2011-02-01
影响因子：
5.1
作者：
Geng, Xuehui;Lou, Haiyan;Drain, Peter
通讯作者：
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影响因子：
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作者：
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作者：
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影响因子：
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