Division of Gene Regulation The Netherlands Cancer Institute, Amsterdam, The Netherlands.
PLoS One. 2011;6(10):e25449. doi: 10.1371/journal.pone.0025449. Epub 2011 Oct 5.
Translocations involving the Mixed Lineage Leukemia (MLL) gene generate in-frame fusions of MLL with more than 50 different partner genes (PGs). Common to all MLL translocations is the exchange not only of coding regions, but also of MLL and PG 3'-untranslated regions (3'UTRs). As a result, the MLL-PG fusion is normally highly expressed and considered the main driver of leukemia development, whereas the function of the PG-MLL fusions in leukemic disease is unclear. As 3'UTRs have been recognized as determinant regions for regulation of gene expression, we hypothesized that loss of the MLL 3'UTR could have a role in generating high MLL-PG levels and leukemia development. Here, we first tested the MLL-PG and PG-MLL mRNA levels in different leukemic cells and tumours and uncovered differential expression that indicates strong repression by the MLL-3'UTR. Reporter assays confirmed that the 3'UTR of MLL, but not of its main PGs, harbours a region that imposes a strong gene silencing effect. Gene suppression by the MLL 3'UTR was largely microRNA independent and did not affect mRNA stability, but inhibited transcription. This effect can at least partially be attributed to a tighter interaction of the MLL 3'UTR with RNA polymerase II than PG 3'UTRs, affecting its phosphorylation state. Altogether, our findings indicate that MLL translocations relieve oncogenic MLL-PG fusions from the repressive MLL 3'UTR, contributing to higher activity of these genes and leukaemia development.
易位涉及混合谱系白血病(MLL)基因,导致 MLL 与 50 多个不同的伙伴基因(PG)形成框内融合。所有 MLL 易位的共同点不仅是编码区的交换,还有 MLL 和 PG 3'非翻译区(3'UTR)的交换。结果,MLL-PG 融合通常高度表达,并被认为是白血病发展的主要驱动因素,而 PG-MLL 融合在白血病中的功能尚不清楚。由于 3'UTR 已被认为是调节基因表达的决定区域,我们假设 MLL 3'UTR 的缺失可能在产生高 MLL-PG 水平和白血病发展中发挥作用。在这里,我们首先在不同的白血病细胞和肿瘤中测试了 MLL-PG 和 PG-MLL mRNA 水平,发现了差异表达,表明 MLL-3'UTR 有很强的抑制作用。报告基因分析证实,MLL 的 3'UTR,而不是其主要 PGs 的 3'UTR,具有一个可施加强烈基因沉默效应的区域。MLL 3'UTR 的基因抑制在很大程度上不依赖 microRNA,也不影响 mRNA 稳定性,但抑制转录。这种效应至少部分归因于 MLL 3'UTR 与 RNA 聚合酶 II 的紧密相互作用,影响其磷酸化状态。总之,我们的研究结果表明,MLL 易位使致癌性 MLL-PG 融合从抑制性 MLL 3'UTR 中解脱出来,有助于这些基因的更高活性和白血病的发展。