A critical role of the Thy28-MYH9 axis in B cell-specific expression of the Pax5 gene in chicken B cells.
A critical role of the Thy28-MYH9 axis in B cell-specific expression of the Pax5 gene in chicken B cells.
复制标题
DOI：
10.1371/journal.pone.0116579
复制
发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Fujii H
中科院分区：
文献类型：
--
作者：
Fujita T;Kitaura F;Fujii H
越来越多的证据表明Pax5在B细胞谱系承诺中起重要作用。然而，Pax5在B细胞特异性表达的分子机制尚不完全清楚。在这里，我们应用插入染色质免疫沉淀（iChIP）结合使用细胞培养氨基酸（SILAC）的稳定同位素标记（iChIP-SILAC）来直接鉴定与内源性单拷贝鸡Pax5基因启动子区域相互作用的蛋白质。通过比较B细胞与巨噬细胞样细胞通过异位表达C/EBPβ进行反分化，iChIP-SILAC检测到B细胞特异性核蛋白Thy28/Thyn1与Pax5 1A启动子的相互作用。B细胞向巨噬细胞样细胞的反分化导致Thy28表达下调。Thy28的功能丧失导致Pax5表达减少和MYH9 （Thy28相互作用蛋白之一）募集到Pax5 1A启动子。MYH9的功能丧失也会导致Pax5表达的降低。因此，我们的分析表明，通过MYH9在鸡B细胞中募集Pax5位点，Thy28在功能上是Pax5在B细胞特异性表达所必需的。
Accumulating evidence suggests that Pax5 plays essential roles in B cell lineage commitment. However, molecular mechanisms of B cell-specific expression of Pax5 are not fully understood. Here, we applied insertional chromatin immunoprecipitation (iChIP) combined with stable isotope labeling using amino acids in cell culture (SILAC) (iChIP-SILAC) to direct identification of proteins interacting with the promoter region of the endogenous single-copy chicken Pax5 gene. By comparing B cells with macrophage-like cells trans-differentiated by ectopic expression of C/EBPβ, iChIP-SILAC detected B cell-specific interaction of a nuclear protein, Thy28/Thyn1, with the Pax5 1A promoter. Trans-differentiation of B cells into macrophage-like cells caused down-regulation of Thy28 expression. Loss-of-function of Thy28 induced decrease in Pax5 expression and recruitment of myosin-9 (MYH9), one of Thy28-interacting proteins, to the Pax5 1A promoter. Loss-of-function of MYH9 also induced decrease in Pax5 expression. Thus, our analysis revealed that Thy28 is functionally required for B cell-specific expression of Pax5 via recruitment of MYH9 to the Pax5 locus in chicken B cells.
登录
查看更多内容
影响因子：
14.9
作者：
Fujita T;Ryser S;Tortola S;Piuz I;Schlegel W
通讯作者：
Schlegel W
影响因子：
7.2
作者：
Compton, MM;Thomson, JM;Icard, AH
通讯作者：
Icard, AH
影响因子：
2.8
作者：
Fujita, Toshitsugu;Fujii, Hodaka
通讯作者：
Fujii, Hodaka
影响因子：
32.4
作者：
Decker, Thorsten;di Magliano, Marina Pasca;Busslinger, Meinrad
通讯作者：
Busslinger, Meinrad
影响因子：
4.4
作者：
Hirano, M;Kikuchi, Y;Takatsu, K
通讯作者：
Takatsu, K