Human endogenous retrovirus K (HML-2) RNA and protein expression is a marker for human embryonic and induced pluripotent stem cells.
Human endogenous retrovirus K (HML-2) RNA and protein expression is a marker for human embryonic and induced pluripotent stem cells.
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DOI：
10.1186/1742-4690-10-115
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发表时间：
2013-10-24
期刊：
影响因子：
3.3
通讯作者：
Löwer R
中科院分区：
文献类型：
--
作者：
Fuchs NV;Loewer S;Daley GQ;Izsvák Z;Löwer J;Löwer R
恶性人类胚胎癌细胞（ECCs）依赖与非恶性胚胎干细胞（ESCs）相似的转录网络来控制自我更新、维持多能性和抑制分化。由于沉默的HERV-K（HML-2）基因座的再激活是ECC的标志，我们询问该HERV组是否也在ESC和诱导多能干细胞（iPSC）中被再激活。使用RT-PCR和Western Blot，我们证明了HERV-K（HML-2）RNA和蛋白在未分化的人ESC和iPSC中的表达。通过胚状体形成诱导分化导致HERV-K（HML-2）前病毒表达的快速沉默。对gag基因保守区的测序分析表明，ESC和iPSC中的前病毒表达代表了66个几乎全长HERV-K（HML-2）基因座中的至少11个，在各个细胞系中具有略微不同的模式。这些前病毒是人类特异性整合的，并且具有启动子活性的长末端重复序列（LTR 5 hs亚组）。我们观察到在所有测试的ECC、ESC和iPSC系中NP 9和Gag编码前病毒K101（22q11.21）和K10（5q33.3）的高mRNA水平，而K37（11q23.3）mRNA仅在ESC和iPSC中检测到。此外，我们在所有研究的细胞系中检测到编码RNA输出接头Rec的前病毒mRNA的表达。来源于K108（7p22.1）基因座的前病毒mRNA，其阿利亚编码功能性Rec和Env蛋白，仅在恶性ECC系中被再激活，而不在良性ESC或iPSC中被再激活。HERV-K（HML-2）RNA和蛋白质表达是多能人类干细胞的标志物。分化的启动导致快速下调。需要进一步的研究来探索HERV-K（HML-2）RNA和蛋白质在多能干细胞中的假定功能作用。
Malignant human embryonal carcinoma cells (ECCs) rely on similar transcriptional networks as non-malignant embryonic stem cells (ESCs) to control selfrenewal, maintain pluripotency, and inhibit differentiation. Because re-activation of silenced HERV-K(HML-2) loci is a hallmark of ECCs, we asked if this HERV group was also reactivated in ESCs and induced pluripotent stem cells (iPSCs). Using RT-PCR and Western Blot, we demonstrate HERV-K(HML-2) RNA and protein expression in undifferentiated human ESCs and iPSCs. Induction of differentiation by embryoid body formation resulted in rapid silencing of HERV-K(HML-2) provirus expression. Sequencing analysis of a conserved region of the gag gene showed that proviral expression in ESCs and iPSCs represents at least 11 of the 66 nearly full length HERV-K(HML-2) loci, with slightly varying patterns in individual cell lines. These proviruses are human specific integrations and harbor promoter competent long terminal repeats (LTR5hs subgroup). We observed high mRNA levels of the NP9 and Gag encoding proviruses K101(22q11.21) in all and K10(5q33.3) in most of the ECC, ESC, and iPSC lines tested, while K37(11q23.3) mRNA was detected only in ESCs and iPSCs. In addition, we detected expression of proviral mRNA encoding the RNA export adaptor Rec in all cell lines studied. Proviral mRNA originating from the K108(7p22.1) locus, which inter alia codes for functional Rec and Env proteins, was only reactivated in malignant ECC lines, not in benign ESCs or iPSCs. HERV-K(HML-2) RNA and protein expression is a marker for pluripotent human stem cells. Initiation of differentiation results in rapid down-regulation. Further studies are needed to explore a putative functional role of HERV-K(HML-2) RNA and proteins in pluripotent stem cells.
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影响因子：
23.9
作者：
Singh AM;Dalton S
通讯作者：
Dalton S
影响因子：
8
作者：
Boese, A;Sauter, M;Mueller-Lantzsch, N
通讯作者：
Mueller-Lantzsch, N
影响因子：
3.3
作者：
Schmitt K;Reichrath J;Roesch A;Meese E;Mayer J
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Mayer J
影响因子：
3.8
作者：
Kaufmann, Sabine;Sauter, Manes;Mueller-Lantzsch, Nikolaus
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Mueller-Lantzsch, Nikolaus
影响因子：
64.8
作者：
Park, In-Hyun;Zhao, Rui;Daley, George Q.
通讯作者：
Daley, George Q.