FLASH结合早幼粒细胞白血病蛋白Ⅳ并增强p53的SUMO修饰
FLASH Interacts with Promyelocytic Leukemia Protein Ⅳ(PML Ⅳ)and Enhances the SUMOylation of p53
王梦妮 1熊真真 1王之盈 2吴建华 1石晓钟2
作者信息
- 1. 华南理工大学生物科学与工程学院生物技术系,广州51006
- 2. 南昌大学医学部基础医学院生理系,南昌 330031
- 折叠
摘要
FLASH/CASP8AP2为基因组中一个独特的基因,参与多个细胞调控过程,包括细胞凋亡、组蛋白基因pre-mRNA的加工、转录调控以及细胞周期进程等.临床医学研究表明,FLASH是急性淋巴细胞白血病的一种预后标志物,还是多种癌细胞存活的关键因子.因此,对FLASH功能的深入研究有望为相关疾病治疗提供新的视角.我们先前鉴定FLASH为p53的结合因子,并发现其能够增强p53的转录活性.在此基础上,本文阐述了 FLASH和p53相互作用的分子机制.研究结果表明,p53 K386 突变显著降低其与 FLASH(aa 51～200)和 FLASH-SIM(aa 1 534～1 806)的结合强度.然而,GST沉降分析仅能检测到SUMO与FLASH-SIM而不是FLASH(aa 51～200)之间的相互作用.在细胞中过量表达FLASH增强了整体性SUMO1修饰以及p53的SUMO1修饰,这可能是FLASH增强p53转录活性的一种作用机制.由于PML NB为细胞内SUMO的亚细胞反应器,而PML Ⅳ亚型能够特异性地增强p53的SUMO修饰,我们在此分析了 FLASH与PML Ⅳ之间的相互作用,并鉴定了二者相互作用的结构域基础:FLASH-N3A(aa 501～802)和FLASH-C2(aa 1 807～1981)均能与PML Ⅳ(aa 228～633)结合.进一步的研究显示,PML Ⅳ能够增强FLASH调控的整体性SUMO修饰和p53的SUMO修饰,即二者在功能上存在协同性.FLASH与肿瘤抑制因子p53和PML Ⅳ之间的相互作用,丰富了对其功能的认识,有望揭示FLASH在肿瘤发生过程中的作用机制,为癌症的诊断和治疗提供新的思路.
Abstract
As a unique gene in the genome,FLASH(FADD-like interleukin-1β-converting enzyme asso-ciated huge protein)/CASP8AP2 is involved in multiple cellular processes,including apoptosis,histone gene pre-mRNA processing,transcriptional regulation,and cell cycle progression.Clinical studies have shown that FLASH is a valuable prognostic marker for acute lymphoblastic leukemia,and a crucial factor for the survival of various cancer cells.Therefore,in-depth research into the function of FLASH may offer new perspectives for the treatment of related diseases.Our previous research identified FLASH as a bind-ing partner of p53,demonstrating that FLASH enhances the transcriptional activity of p53.Here we fur-ther investigate the molecular mechanisms of the interaction between FLASH and p53,revealing that the p53-K386R mutation(SUMOylation residue)attenuated its interaction with FLASH(aa 51-200)and FLASH-SIM(SUMO-interacting motif)(aa 1 534-1 806)significantly.However,SUMO can bind to FLASH-SIM directly,instead of FLASH(aa 51-200).Subsequent research shows that overexpression of FLASH in cells enhances global SUMO1 conjugation and p53-SUMO1 conjugation,therefore providing a plausible explanation for the underlying mechanism of FLASH enhancing the transcriptional activity of p53.Since promyelocytic leukemia protein nuclear body(PML NB)serves as subcellular reactors for SUMO conjugation within the cell,and the PML Ⅳ isoform can specifically enhance the SUMO modifica-tion of p53,we have investigated the interaction between FLASH and PML Ⅳ,and elucidated the struc-tural basis of their interaction:both FLASH-N3A(501-802)and FLASH-C2(1 807-1 981)bind to PML Ⅳ(aa 228-633).Further investigations reveal that they can synergistically enhance global SUMO1 modification as well as SUMO1 modification of p53.The interaction between FLASH and tumor suppres-sors p53 or PML Ⅳ enriches our understanding of its function and reveals the potential mechanism of FLASH in tumor development,therefore offering novel insights into cancer diagnosis and treatment.
关键词
FADD样白细胞介素1β转化酶相关巨蛋白/p53/小泛素样修饰/早幼粒细胞白血病蛋白ⅣKey words
FADD-like interleukin-1β-converting enzyme associated huge protein(FLASH)/p53/SU-MOylation/promyelocytic leukemia protein Ⅳ(PML Ⅳ)引用本文复制引用
出版年
2024
NETL
NSTL
万方数据