5'UTR 序列在 BDNF mRNA 树突贩运中的独特作用:BDNF 剪接变异体空间编码的附加机制。

Distinct role of 5'UTR sequences in dendritic trafficking of BDNF mRNA: additional mechanisms for the BDNF splice variants spatial code.

机构信息

Department of Life Sciences (Q Building), University of Trieste, Via Licio Giorgieri, 5, 34127, Trieste, Italy.

出版信息

Mol Brain. 2021 Jan 12;14(1):10. doi: 10.1186/s13041-020-00680-8.

Abstract

The neurotrophin Brain-derived neurotrophic factor (BDNF) is encoded by multiple bipartite transcripts. Each BDNF transcript is composed by one out of 11 alternatively spliced exons containing the 5'untranslated region (UTR), and one common exon encompassing the coding sequence (CDS) and the 3'UTR with two variants (short and long). In neurons, BDNF mRNA variants have a distinct subcellular distribution, constituting a "spatial code", with exon 1, 3, 5, 7 and 8 located in neuronal somata, exon 4 extending into proximal dendrites, and exon 2 and 6 reaching distal dendrites. We previously showed that the CDS encodes constitutive dendritic targeting signals (DTS) and that both the 3'UTR-short and the 3'UTR-long contain activity-dependent DTS. However, the role of individual 5'UTR exons in mRNA sorting remains unclear. Here, we tested the ability of each different BDNF 5'UTRs to affect the subcellular localization of the green fluorescent protein (GFP) reporter mRNA. We found that exon 2 splicing isoforms (2a, 2b, and 2c) induced a constitutive dendritic targeting of the GFP reporter mRNA towards distal dendritic segments. The other isoforms did not affect GFP-mRNA dendritic trafficking. Through a bioinformatic analysis, we identified five unique cis-elements in exon 2a, 2b, and 2c which might contribute to building a DTS. This study provides additional information on the mechanism regulating the cellular sorting of BDNF mRNA variants.

摘要

神经营养因子脑源性神经营养因子 (BDNF) 由多个二分体转录本编码。每个 BDNF 转录本由一个包含 5'非翻译区 (UTR) 的 11 个选择性剪接外显子之一和一个包含编码序列 (CDS) 和 3'UTR 的共同外显子组成,其中有两种变体(短和长)。在神经元中,BDNF mRNA 变体具有独特的亚细胞分布,构成了一种“空间密码”,外显子 1、3、5、7 和 8 位于神经元胞体中,外显子 4 延伸到近端树突,外显子 2 和 6 到达远端树突。我们之前表明,CDS 编码组成型树突靶向信号 (DTS),并且 3'UTR-短和 3'UTR-长都包含活性依赖性 DTS。然而,个体 5'UTR 外显子在 mRNA 分拣中的作用仍不清楚。在这里,我们测试了每个不同的 BDNF 5'UTRs 影响绿色荧光蛋白 (GFP) 报告 mRNA 亚细胞定位的能力。我们发现外显子 2 剪接异构体 (2a、2b 和 2c) 诱导 GFP 报告 mRNA 向远端树突段的组成型树突靶向。其他异构体不会影响 GFP-mRNA 树突运输。通过生物信息学分析,我们在外显子 2a、2b 和 2c 中鉴定了五个独特的顺式元件,它们可能有助于构建 DTS。这项研究提供了关于调节 BDNF mRNA 变体细胞分拣的机制的更多信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec20/7805101/b018f81858c2/13041_2020_680_Fig1_HTML.jpg

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