DNA crosslinking and recombination-activating genes 1/2 (RAG1/2) are required for oncogenic splicing in acute lymphoblastic leukemia.
DNA crosslinking and recombination-activating genes 1/2 (RAG1/2) are required for oncogenic splicing in acute lymphoblastic leukemia.
复制标题
DOI：
10.1002/cac2.12234
复制
发表时间：
2021-11
期刊：
影响因子：
--
通讯作者：
Meng G
中科院分区：
文献类型：
--
作者：
Zhang H;Cheng N;Li Z;Bai L;Fang C;Li Y;Zhang W;Dong X;Jiang M;Liang Y;Zhang S;Mi J;Zhu J;Zhang Y;Chen SJ;Zhao Y;Weng XQ;Hu W;Chen Z;Huang J;Meng G
异常的选择性剪接常与癌症发生有关。在B-细胞急性淋巴细胞白血病（B-ALL）中，与免疫球蛋白重链融合的双同源框4（DUX 4/IGH）可导致E-26转化特异性家族相关基因异常转录本（ERGalt）和其他剪接变体的异常产生。然而，支持这一过程的分子机制仍然难以捉摸。在这里，我们的目的是了解DUX 4/IGH如何触发白血病的异常剪接。进行差异内含子保留分析以鉴定B‐ALL患者中的新型DUX 4/IGH驱动的剪接。使用X射线晶体学、小角X射线散射（SAXS）和分析超离心来研究DUX 4/IGH如何识别双DUX 4响应元件（DRE）-DRE位点。进行ERGalt生物发生和B-细胞分化测定以表征DUX 4/IGH交联活性。为了检查DUX 4/IGH驱动的剪接是否需要重组激活基因1/2（RAG 1/2），进行了邻位连接试验、免疫共沉淀、哺乳动物双杂交表征、体外RAG 1/2切割和shRNA敲低试验。我们报告了C型凝集素结构域家族12成员A异常转录物（CLEC 12 Aalt）和6号染色体开放阅读框89异常转录物（C6 orf 89 alt）中先前未被识别的内含子保留事件，其中也含有重复的DRE-DRE位点。支持性地，X射线晶体学和SAXS表征揭示DUX 4同源框结构域（HD）1-HD2可能二聚成哑铃形反式构型以交联两个相邻的DRE位点。受损的DUX 4/IGH介导的交联消除了ERGalt、CLEC 12 Aalt和C6 orf 89 alt生物合成，导致其对B细胞分化的抑制作用显著减轻。此外，我们还观察到DUX 4/IGH靶基因中罕见的RAG 1/2介导的重组信号序列样DNA编辑。支持性的是，白血病Reh细胞中RAG 12的shRNA敲低始终损害ERGalt，CLEC 12 Aalt和C6 orf 89 alt的生物合成。所有这些结果表明，DUX 4/IGH驱动的DNA交联是RAG 1/2募集到双串联DRE-DRE位点所必需的，催化急性淋巴细胞白血病中的V（D）J样重组和致癌剪接。
Abnormal alternative splicing is frequently associated with carcinogenesis. In B‐cell acute lymphoblastic leukemia (B‐ALL), double homeobox 4 fused with immunoglobulin heavy chain (DUX4/IGH) can lead to the aberrant production of E‐26 transformation‐specific family related gene abnormal transcript (ERGalt) and other splicing variants. However, the molecular mechanism underpinning this process remains elusive. Here, we aimed to know how DUX4/IGH triggers abnormal splicing in leukemia. The differential intron retention analysis was conducted to identify novel DUX4/IGH‐driven splicing in B‐ALL patients. X‐ray crystallography, small angle X‐ray scattering (SAXS), and analytical ultracentrifugation were used to investigate how DUX4/IGH recognize double DUX4 responsive element (DRE)‐DRE sites. The ERGalt biogenesis and B‐cell differentiation assays were performed to characterize the DUX4/IGH crosslinking activity. To check whether recombination‐activating gene 1/2 (RAG1/2) was required for DUX4/IGH‐driven splicing, the proximity ligation assay, co‐immunoprecipitation, mammalian two hybrid characterizations, in vitro RAG1/2 cleavage, and shRNA knock‐down assays were performed. We reported previously unrecognized intron retention events in C‐type lectin domain family 12, member A abnormal transcript (CLEC12Aalt) and chromosome 6 open reading frame 89 abnormal transcript (C6orf89alt), where also harbored repetitive DRE‐DRE sites. Supportively, X‐ray crystallography and SAXS characterization revealed that DUX4 homeobox domain (HD)1‐HD2 might dimerize into a dumbbell‐shape trans configuration to crosslink two adjacent DRE sites. Impaired DUX4/IGH‐mediated crosslinking abolishes ERGalt, CLEC12Aalt, and C6orf89alt biogenesis, resulting in marked alleviation of its inhibitory effect on B‐cell differentiation. Furthermore, we also observed a rare RAG1/2‐mediated recombination signal sequence‐like DNA edition in DUX4/IGH target genes. Supportively, shRNA knock‐down of RAG1/2 in leukemic Reh cells consistently impaired the biogenesis of ERGalt, CLEC12Aalt, and C6orf89alt. All these results suggest that DUX4/IGH‐driven DNA crosslinking is required for RAG1/2 recruitment onto the double tandem DRE‐DRE sites, catalyzing V(D)J‐like recombination and oncogenic splicing in acute lymphoblastic leukemia.
登录
查看更多内容
小儿急性淋巴细胞白血病中的染色质可及性鉴定亚型特异性染色质景观和基因调节网络。
影响因子：
11.4
作者：
Diedrich JD;Dong Q;Ferguson DC;Bergeron BP;Autry RJ;Qian M;Yang W;Smith C;Papizan JB;Connelly JP;Hagiwara K;Crews KR;Pruett-Miller SM;Pui CH;Yang JJ;Relling MV;Evans WE;Savic D
通讯作者：
Savic D
影响因子：
30.8
作者：
Holmfeldt, Linda;Wei, Lei;Diaz-Flores, Ernesto;Walsh, Michael;Zhang, Jinghui;Ding, Li;Payne-Turner, Debbie;Churchman, Michelle;Andersson, Anna;Chen, Shann-Ching;McCastlain, Kelly;Becksfort, Jared;Ma, Jing;Wu, Gang;Patel, Samir N.;Heatley, Susan L.;Phillips, Letha A.;Song, Guangchun;Easton, John;Parker, Matthew;Chen, Xiang;Rusch, Michael;Boggs, Kristy;Vadodaria, Bhavin;Hedlund, Erin;Drenberg, Christina;Baker, Sharyn;Pei, Deqing;Cheng, Cheng;Huether, Robert;Lu, Charles;Fulton, Robert S.;Fulton, Lucinda L.;Tabib, Yashodhan;Dooling, David J.;Ochoa, Kerri;Minden, Mark;Lewis, Ian D.;To, L. Bik;Marlton, Paula;Roberts, Andrew W.;Raca, Gordana;Stock, Wendy;Neale, Geoffrey;Drexler, Hans G.;Dickins, Ross A.;Ellison, David W.;Shurtleff, Sheila A.;Pui, Ching-Hon;Ribeiro, Raul C.;Devidas, Meenakshi;Carroll, Andrew J.;Heerema, Nyla A.;Wood, Brent;Borowitz, Michael J.;Gastier-Foster, Julie M.;Raimondi, Susana C.;Mardis, Elaine R.;Wilson, Richard K.;Downing, James R.;Hunger, Stephen P.;Loh, Mignon L.;Mullighan, Charles G.
通讯作者：
Mullighan, Charles G.
影响因子：
16.6
作者：
Gu, Zhaohui;Churchman, Michelle;Roberts, Kathryn;Li, Yongjin;Liu, Yu;Harvey, Richard C.;McCastlain, Kelly;Reshmi, Shalini C.;Payne-Turner, Debbie;Iacobucci, Ilaria;Shao, Ying;Chen, I-Ming;Valentine, Marcus;Pei, Deqing;Mungall, Karen L.;Mungall, Andrew J.;Ma, Yussanne;Moore, Richard;Marra, Marco;Stonerock, Eileen;Gastier-Foster, Julie M.;Devidas, Meenakshi;Dai, Yunfeng;Wood, Brent;Borowitz, Michael;Larsen, Eric E.;Maloney, Kelly;Mattano, Leonard A., Jr.;Angiolillo, Anne;Salzer, Wanda L.;Burke, Michael J.;Gianni, Francesca;Spinelli, Orietta;Radich, Jerald P.;Minden, Mark D.;Moorman, Anthony V.;Patel, Bella;Fielding, Adele K.;Rowe, Jacob M.;Luger, Selina M.;Bhatia, Ravi;Aldoss, Ibrahim;Forman, Stephen J.;Kohlschmidt, Jessica;Mrozek, Krzysztof;Marcucci, Guido;Bloomfield, Clara D.;Stock, Wendy;Kornblau, Steven;Kantarjian, Hagop M.;Konopleva, Marina;Paietta, Elisabeth;Willman, Cheryl L.;Loh, Mignon L.;Hunger, Stephen P.;Mullighan, Charles G.
通讯作者：
Mullighan, Charles G.
影响因子：
20.3
作者：
Harvey, Richard C.;Mullighan, Charles G.;Willman, Cheryl L.
通讯作者：
Willman, Cheryl L.
影响因子：
2.9
作者：
Abdiche, Yasmina;Malashock, Dan;Pons, Jaurne
通讯作者：
Pons, Jaurne