Requirement of PML SUMO interacting motif for RNF4- or arsenic trioxide-induced degradation of nuclear PML isoforms.
Requirement of PML SUMO interacting motif for RNF4- or arsenic trioxide-induced degradation of nuclear PML isoforms.
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DOI：
10.1371/journal.pone.0044949
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Chelbi-Alix MK
中科院分区：
文献类型：
--
作者：
Maroui MA;Kheddache-Atmane S;El Asmi F;Dianoux L;Aubry M;Chelbi-Alix MK
PML是核小体（NB）的组织者，以几种命名为PMLI至VII的同种型表达，由于单个基因的选择性剪接，这些同种型的C-末端区域不同。这种变异性对于不同PML亚型的功能很重要。PML NB的形成需要SUMO与PML的共价连接。三氧化二砷（As 2 O3）增强PML SUMO化，导致PML NB大小增加，并促进其与RNF 4（一种负责蛋白酶体介导的PML降解的聚SUMO依赖性泛素E3连接酶）的相互作用。此外，在PML的C-末端区域内存在真正的SUMO相互作用基序（SIM）似乎是在PML NB内募集其他SUMO化蛋白所必需的。该基序存在于所有PML亚型中，除了核PMLVI和胞质PMLVII。使用生物发光共振能量转移（BRET）测定在活细胞中，我们发现，As 2 O3增强SUMO化和核PML亚型（I至VI）与RNF 4的相互作用。此外，在核PML亚型，只有一个缺乏SIM序列，PMLVI，是抗As 2 O3诱导的PML降解。同样，突变的SIM在PMLIII废除其敏感性As 2 O3诱导的降解。PMLVI和PMLIII-SIM突变体仍与RNF 4相互作用。然而，它们对降解过程的抗性是由于它们不能被多聚泛素化，不能有效地募集PML NB中蛋白酶体的20 S核心和11 S复合物的β调节亚基。RNF 4的过表达减轻了PMLVI对As 2 O3诱导的降解的这种抗性。我们的研究结果表明，SIM的PML是不稳定的PML SUMO化和与RNF 4的相互作用，但需要有效的PML泛素化，招聘的NB内的蛋白酶体成分和蛋白酶体依赖性降解的PML响应As 2 O3。
PML, the organizer of nuclear bodies (NBs), is expressed in several isoforms designated PMLI to VII which differ in their C-terminal region due to alternative splicing of a single gene. This variability is important for the function of the different PML isoforms. PML NB formation requires the covalent linkage of SUMO to PML. Arsenic trioxide (As2O3) enhances PML SUMOylation leading to an increase in PML NB size and promotes its interaction with RNF4, a poly-SUMO-dependent ubiquitin E3 ligase responsible for proteasome-mediated PML degradation. Furthermore, the presence of a bona fide SUMO Interacting Motif (SIM) within the C-terminal region of PML seems to be required for recruitment of other SUMOylated proteins within PML NBs. This motif is present in all PML isoforms, except in the nuclear PMLVI and in the cytoplasmic PMLVII. Using a bioluminescence resonance energy transfer (BRET) assay in living cells, we found that As2O3 enhanced the SUMOylation and interaction with RNF4 of nuclear PML isoforms (I to VI). In addition, among the nuclear PML isoforms, only the one lacking the SIM sequence, PMLVI, was resistant to As2O3-induced PML degradation. Similarly, mutation of the SIM in PMLIII abrogated its sensitivity to As2O3-induced degradation. PMLVI and PMLIII-SIM mutant still interacted with RNF4. However, their resistance to the degradation process was due to their inability to be polyubiquitinated and to recruit efficiently the 20S core and the β regulatory subunit of the 11S complex of the proteasome in PML NBs. Such resistance of PMLVI to As2O3-induced degradation was alleviated by overexpression of RNF4. Our results demonstrate that the SIM of PML is dispensable for PML SUMOylation and interaction with RNF4 but is required for efficient PML ubiquitination, recruitment of proteasome components within NBs and proteasome-dependent degradation of PML in response to As2O3.
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影响因子：
3.3
作者：
Geoffroy MC;Jaffray EG;Walker KJ;Hay RT
通讯作者：
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影响因子：
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作者：
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50.3
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通讯作者：
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