CST Is Epistatic With Shieldin to Limit DNA Double-Strand Break End Resection and Promote Repair During Igh Class Switch Recombination

IF 4.5 3区 医学 Q2 IMMUNOLOGY
Chloé Lescale, Timea Marton, Amaury Vaysse, Guillaume Rode, Estelle Vincendeau, Alice Libri, François Dossin, Ludovic Deriano
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引用次数: 0

Abstract

Downstream of 53BP1-RIF1 lies the Shieldin (SHLD) protein complex, which comprises MAD2L2/REV7, SHLD3, SHLD2, and SHLD1, and the CTC1-STN1-TEN1 (CST) complex. During immunoglobulin heavy-chain (Igh) class switch recombination (CSR), 53BP1-RIF1-SHLD promotes productive end-joining by limiting resection of activation-induced cytidine deaminase (AID)-generated DNA double-strand break (DSB) ends. The precise role of the CST complex and its interplay with SHLD during CSR is however elusive. Here, we established AID-inducible B cell lines deficient for CTC1, SHLD1, or both and analyzed CSR in these cells. We show that stimulated CTC1-deficient B cells are defective for IgM-to-IgA class switching, accumulate Igh chromosome breaks and translocations, and display increased end-resection and micro-homology usage at switching sites, demonstrating that CTC1 is essential to suppress alternative end-joining during CSR. We show that CTC1 and SHLD1 are epistatic in preventing exacerbated DNA end resection and genetic instability during CSR. Moreover, using a complementation approach in Shld1 knockout splenic B cells, we show that a SHLD1 mutant defective in CST binding (SHLD1ΔLDLP) is fully proficient for IgM-to-IgG1, IgG2b, IgG3, and IgA class switching, thus demonstrating that the SHLD1-CTC1 interaction through this motif is dispensable for CST and SHLD functions in promoting CSR.

Abstract Image

CST与屏蔽蛋白的上位性作用限制DNA双链断裂端切除,促进DNA修复
53BP1-RIF1的下游是屏蔽蛋白(SHLD)复合物,包括MAD2L2/REV7、SHLD3、SHLD2和SHLD1,以及CTC1-STN1-TEN1 (CST)复合物。在免疫球蛋白重链(Igh)类开关重组(CSR)过程中,53BP1-RIF1-SHLD通过限制激活诱导胞苷脱氨酶(AID)产生的DNA双链断裂(DSB)末端的切除来促进生产端连接。然而,在CSR过程中,CST复合物的确切作用及其与SHLD的相互作用尚不清楚。在这里,我们建立了艾滋病诱导的缺乏CTC1、SHLD1或两者的B细胞系,并分析了这些细胞的CSR。我们发现受刺激的缺乏CTC1的B细胞在igm到iga类转换中存在缺陷,积累了Igh染色体断裂和易位,并且在开关位点显示出增加的末端切除和微同源性使用,表明CTC1对抑制CSR期间的替代末端连接至关重要。我们发现CTC1和SHLD1在防止CSR期间加剧的DNA末端切除和遗传不稳定性方面具有上位性。此外,通过在Shld1敲除的脾B细胞中使用互补方法,我们发现在CST结合上有缺陷的Shld1突变体(SHLD1ΔLDLP)完全精通igm到igg1、IgG2b、IgG3和IgA类转换,从而证明通过该基序的Shld1 - ctc1相互作用对于CST和SHLD促进CSR的功能是必不可少的。
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来源期刊
CiteScore
8.30
自引率
3.70%
发文量
224
审稿时长
2 months
期刊介绍: The European Journal of Immunology (EJI) is an official journal of EFIS. Established in 1971, EJI continues to serve the needs of the global immunology community covering basic, translational and clinical research, ranging from adaptive and innate immunity through to vaccines and immunotherapy, cancer, autoimmunity, allergy and more. Mechanistic insights and thought-provoking immunological findings are of interest, as are studies using the latest omics technologies. We offer fast track review for competitive situations, including recently scooped papers, format free submission, transparent and fair peer review and more as detailed in our policies.
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