Mutations within the LINC-HELLP non-coding RNA differentially bind ribosomal and RNA splicing complexes and negatively affect trophoblast differentiation
Mutations within the LINC-HELLP non-coding RNA differentially bind ribosomal and RNA splicing complexes and negatively affect trophoblast differentiation
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DOI：
10.1093/hmg/ddv274
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发表时间：
2015-10-01
影响因子：
3.5
通讯作者：
Oudejans, Cees B. M.
中科院分区：
文献类型：
--
作者：
van Dijk, Marie;Visser, Allerdien;Oudejans, Cees B. M.
在荷兰家族中，LINC-HELLP显示了与妊娠特异性HELLP综合征的染色体连锁，减少了妊娠早期绒毛外滋养层细胞从增殖型到侵袭型的分化。在这里，我们发现在HELLP家族中发现的LINC-HELLP突变通过诱导增殖速度或通过导致细胞周期退出而对滋养层细胞的分化产生负面影响，表现为增殖和侵袭的减少。由于lincRNAs主要通过与蛋白质的相互作用发挥作用，我们使用染色质分离、RNA纯化和蛋白质质谱来鉴定直接相互作用的蛋白质。我们发现22个蛋白质主要聚集在两个功能网络中，即RNA剪接和核糖体。验证了YBX1、PCBP1、PCBP2、RPS6和RPL7与这些蛋白的结合受所携带的HELLP突变的影响。最后，我们发现LINC-HELLP转录水平在怀孕前三个月的妇女中显著上调，而在后来出现妊娠并发症的试验组患者中似乎没有这种上调，这表明了它在体内的功能意义。
LINC-HELLP, showing chromosomal linkage with the pregnancy-specific HELLP syndrome in Dutch families, reduces differentiation from a proliferative to an invasive phenotype of first-trimester extravillous trophoblasts. Here we show that mutations in LINC-HELLP identified in HELLP families negatively affect this trophoblast differentiation either by inducing proliferation rate or by causing cell cycle exit as shown by a decrease in both proliferation and invasion. As LincRNAs predominantly function through interactions with proteins, we identified the directly interacting proteins using chromatin isolation by RNA purification followed by protein mass spectrometry. We found 22 proteins predominantly clustering in two functional networks, i.e. RNA splicing and the ribosome. YBX1, PCBP1, PCBP2, RPS6 and RPL7 were validated, and binding to these proteins was influenced by the HELLP mutations carried. Finally, we show that the LINC-HELLP transcript levels are significantly upregulated in plasma of women in their first trimester of pregnancy compared with non-pregnant women, whereas this upregulation seems absent in a pilot set of patients later developing pregnancy complications, indicative of its functional significance in vivo.