The Prp19/Pso4 core complex undergoes ubiquitylation and structural alterations in response to DNA damage
The Prp19/Pso4 core complex undergoes ubiquitylation and structural alterations in response to DNA damage
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DOI：
10.1016/j.bbrc.2007.01.097
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发表时间：
2007-03-23
影响因子：
3.1
通讯作者：
Legerski, Randy J.
中科院分区：
文献类型：
--
作者：
Lu, Xiaoyan;Legerski, Randy J.
Prp19/Pso4 是一种含有 E3 连接酶的 U 型盒，在前 mRNA 剪接中具有已被证明的作用，但在酵母和哺乳动物细胞中也被认为在 DNA 损伤处理中具有直接作用。在本报告中，我们提供了进一步的证据来支持后一种说法。我们发现 hPrp19 形成泛素化寡聚体，在非还原条件下能够抵抗 SDS 凝胶电泳的破坏，表明这是由泛素和 Prp19 中的半胱氨酸残基之间的硫醇酯介导的。用 DNA 损伤剂处理细胞后，该物种的水平显着增强，并且其与染色质的关联增加。此外，已知 hPrp19 与 Cdc5L、Plrg1 和 Spf27 形成稳定的核心复合物；然而，泛素化的 hPrp19 无法与 Cdc5L 或 Plrg1 相互作用，这表明 DNA 损伤可以诱导 hPrp19 核心复合物发生深刻的改变。最后，我们发现人类细胞中 hPrp19 的过度表达具有促生存作用，因为它降低了细胞暴露于 DNA 损伤后观察到的细胞凋亡水平。 (c) 2067 Elsevier Inc. 保留所有权利。
Prp19/Pso4, a U-box containing E3 ligase, has a demonstrated role in pre-mRNA splicing, but has also been implicated in both yeast and mammalian cells as having a direct role in DNA damage processing. In this report, we provide further evidence in support of this latter assertion. We show that hPrp19 forms an ubiquitylated oligomeric species that is resistant to disruption by SDS gel electrophoresis under nonreducing conditions suggesting that is mediated by a thiolester between ubiquitin and a cysteine residue in Prp19. The level of this species is significantly enhanced upon treatment of cells with DNA damaging agents, and its association with chromatin is increased. In addition, hPrp19 is known to form a stable core complex with Cdc5L, Plrg1, and Spf27; however, ubiquitylated hPrp19 fails to interact with either Cdc5L or Plrg1 indicating that DNA damage can induce profound alterations to the hPrp19 core complex. Finally, we show that overexpression of hPrp19 in human cells provides a pro-survival affect in that it reduces the levels of apoptosis observed after exposure of cells to DNA damage. (c) 2067 Elsevier Inc. All rights reserved.