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基因组和 cDNA 选择扩增鉴定了果蝇蛋白性致死的转录组范围结合位点。

Genomic and cDNA selection-amplification identifies transcriptome-wide binding sites for the Drosophila protein sex-lethal.

机构信息

Department of Molecular, Cellular and Developmental Biology, University of Colorado at Boulder, Boulder, CO, United States of America.

出版信息

PLoS One. 2021 May 24;16(5):e0250592. doi: 10.1371/journal.pone.0250592. eCollection 2021.

Abstract

BACKGROUND

Downstream targets for a large number of RNA-binding proteins remain to be identified. The Drosophila master sex-switch protein Sex-lethal (SXL) is an RNA-binding protein that controls splicing, polyadenylation, or translation of certain mRNAs to mediate female-specific sexual differentiation. Whereas some targets of SXL are known, previous studies indicate that additional targets of SXL have escaped genetic screens.

METHODOLOGY/PRINCIPAL FINDINGS: Here, we have used an alternative molecular approach of GEnomic Selective Enrichment of Ligands by Exponential enrichment (GESELEX) using both the genomic DNA and cDNA pools from several Drosophila developmental stages to identify new potential targets of SXL. Our systematic analysis provides a comprehensive view of the Drosophila transcriptome for potential SXL-binding sites.

CONCLUSION/SIGNIFICANCE: We have successfully identified new SXL-binding sites in the Drosophila transcriptome. We discuss the significance of our analysis and that the newly identified binding sites and sequences could serve as a useful resource for the research community. This approach should also be applicable to other RNA-binding proteins for which downstream targets are unknown.

摘要

背景

大量 RNA 结合蛋白的下游靶标仍有待鉴定。果蝇主要性别转换蛋白 Sex-lethal(SXL)是一种 RNA 结合蛋白,可控制某些 mRNA 的剪接、多聚腺苷酸化或翻译,从而介导雌性特异性性别分化。尽管已经知道 SXL 的一些靶标,但以前的研究表明,SXL 的其他靶标已经逃脱了遗传筛选。

方法/主要发现:在这里,我们使用了一种替代的分子方法,即通过指数富集的基因组选择性配体富集(GESELEX),使用来自几个果蝇发育阶段的基因组 DNA 和 cDNA 池,来鉴定 SXL 的新潜在靶标。我们的系统分析为果蝇转录组中 SXL 结合位点提供了全面的视图。

结论/意义:我们已经成功地在果蝇转录组中鉴定了新的 SXL 结合位点。我们讨论了我们分析的意义,并且新鉴定的结合位点和序列可以作为研究社区的有用资源。这种方法也应该适用于其他下游靶标未知的 RNA 结合蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9064/8143406/507499f461ac/pone.0250592.g001.jpg

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