The role of transcriptional corepressor Nif3l1 in early stage of neural differentiation via cooperation with Trip15/CSN2
The role of transcriptional corepressor Nif3l1 in early stage of neural differentiation via cooperation with Trip15/CSN2
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DOI：
10.1074/jbc.m209856200
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发表时间：
2003-03-21
影响因子：
4.8
通讯作者：
Tashiro, F
中科院分区：
文献类型：
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作者：
Akiyama, H;Fujisawa, N;Tashiro, F
小鼠Nif3l1基因从细菌到人类高度保守。尽管该基因在胚胎发育过程中表达，但其生物学功能仍不清楚。在这里，我们发现Nif3l1通过与COP9信号体的转录辅抑制因子/组分Trip15/CSN2合作，参与了视黄酸引发的P19胚胎癌细胞的神经分化。以Trip15/CSN2为诱饵，通过酵母双杂交筛选，从P19细胞cDNA文库中分离到Nif3l1 cDNA。这种相互作用通过下拉试验和表位标记的共免疫沉淀得到证实。虽然Nif3l1主要存在于细胞质中，但在视黄酸诱导的P19细胞神经分化过程中，也观察到Nif3l1向细胞核中的易位，并通过Trip15/CSN2的强制表达而增强。此外，强化Nif311义RNA而非反义RNA的表达，可促进P19细胞的神经分化，并伴有Oct-3/4 mRNA表达的强烈下调和msh -1 mRNA表达的快速诱导。荧光素酶报告基因实验表明，Nif3l1可以作为转录抑制因子，并协同Trip15/CSN2的转录抑制。这些结果表明，Nif3l1通过与Trip15/CSN2的合作参与神经分化。
Mouse Nif3l1 gene is highly conserved from bacteria to human. Even though this gene is expressed throughout embryonic development, its biological - function is still obscure. Here, we show that Nif3l1 participates in retinoic acid-primed neural differentiation of P19 embryonic carcinoma cells through cooperation with Trip15/CSN2, a transcriptional corepressor/component of COP9 signalosome. We isolated Nif3l1 cDNA from P19 cell cDNA library by a yeast two-hybrid screening using Trip15/CSN2 as a bait. This interaction was confirmed by a pull-down assay and an epitope-tagged coimmunoprecipitation. Although Nif3l1 was mainly detected in the cytoplasm, the translocation of Nif3l1 into the nuclei was observed in retinoic acid-primed neural differentiation of P19 cells and enhanced by the enforced expression of Trip15/CSN2. Furthermore, enforced expression of sense Nif311 RNA, but not antisense RNA, enhanced the neural differentiation of P19 cells accompanying the intense down-regulation of Oct-3/4 mRNA expression and the rapid induction of Mash-1 mRNA expression. Luciferase reporter assay showed that Nif3l1 could act as a transcriptional repressor and synergized the transcriptional repression by Trip15/CSN2. These results indicate that Nif3l1 implicates in neural differentiation through the cooperation with Trip15/CSN2.