鸡 miR-150 靶向功能斑马鱼 MYB 3'UTR 靶位点的禽类同源物。

The chicken miR-150 targets the avian orthologue of the functional zebrafish MYB 3'UTR target site.

机构信息

Université François Rabelais, Equipe Transcription, Lymphome Viro-induit, UFR Sciences et Techniques, Parc de Grandmont, F-37200 Tours, France.

出版信息

BMC Mol Biol. 2010 Sep 2;11:67. doi: 10.1186/1471-2199-11-67.

DOI:10.1186/1471-2199-11-67

PMID:20813039

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2940766/

Abstract

BACKGROUND

The c-myb proto-oncogene is the founding member of a family of transcription factors involved principally in haematopoiesis, in diverse organisms, from zebrafish to mammals. Its deregulation has been implicated in human leukaemogenesis and other cancers. The expression of c-myb is tightly regulated by post-transcriptional mechanisms involving microRNAs. MicroRNAs are small, highly conserved non-coding RNAs that inhibit translation and decrease mRNA stability by binding to regulatory motifs mostly located in the 3'UTR of target mRNAs conserved throughout evolution. MYB is an evolutionarily conserved miR-150 target experimentally validated in mice, humans and zebrafish. However, the functional miR-150 sites of humans and mice are orthologous, whereas that of zebrafish is different.

RESULTS

We identified the avian mature miRNA-150-5P, Gallus gallus gga-miR-150 from chicken leukocyte small-RNA libraries and showed that, as expected, the gga-miR-150 sequence was highly conserved, including the seed region sequence present in the other miR-150 sequences listed in miRBase. Reporter assays showed that gga-miR-150 acted on the avian MYB 3'UTR and identified the avian MYB target site involved in gga-miR-150 binding. A comparative in silico analysis of the miR-150 target sites of MYB 3'UTRs from different species led to the identification of a single set of putative target sites in amphibians and zebrafish, whereas two sets of putative target sites were identified in chicken and mammals. However, only the target site present in the chicken MYB 3'UTR that was identical to that in zebrafish was functional, despite the additional presence of mammalian target sites in chicken. This specific miR-150 site usage was not cell-type specific and persisted when the chicken c-myb 3'UTR was used in the cell system to identify mammalian target sites, showing that this miR-150 target site usage was intrinsic to the chicken c-myb 3'UTR.

CONCLUSION

Our study of the avian MYB/gga-miR-150 interaction shows a conservation of miR-150 target site functionality between chicken and zebrafish that does not extend to mammals.

摘要

背景

c-myb 原癌基因是一个参与造血的转录因子家族的创始成员，在从斑马鱼到哺乳动物等不同生物中都有发现。其失调与人类白血病发生和其他癌症有关。c-myb 的表达受涉及 microRNA 的转录后机制的严格调控。microRNA 是小的、高度保守的非编码 RNA，通过结合到进化过程中保守的靶 mRNA 的 3'UTR 中的调节基序，抑制翻译并降低 mRNA 稳定性。MYB 是在小鼠、人类和斑马鱼中实验验证的保守的 miR-150 靶标。然而，人类和小鼠的功能性 miR-150 位点是同源的，而斑马鱼的则不同。

结果

我们从鸡白细胞小 RNA 文库中鉴定出禽类成熟 miRNA-150-5P，即 Gallus gallus gga-miR-150，并表明，正如预期的那样，gga-miR-150 序列高度保守，包括在 miRBase 中列出的其他 miR-150 序列中的种子序列。报告基因实验表明，gga-miR-150 作用于禽类 MYB 3'UTR，并鉴定出与 gga-miR-150 结合相关的禽类 MYB 靶位点。对不同物种 MYB 3'UTR 的 miR-150 靶位点进行比较计算机分析，导致在两栖动物和斑马鱼中鉴定出一组单一的假定靶位点，而在鸡和哺乳动物中鉴定出两组假定靶位点。然而，尽管在鸡中存在哺乳动物靶位点，但只有在鸡 MYB 3'UTR 中与斑马鱼相同的靶位点是功能性的。这种特定的 miR-150 靶位点的使用不是细胞类型特异性的，并且当鸡 c-myb 3'UTR 用于细胞系统鉴定哺乳动物靶位点时仍然存在，表明这种 miR-150 靶位点的使用是鸡 c-myb 3'UTR 固有的。