Regulation of the Signal-Dependent E Protein HEBAlt Through a YYY Motif Is Required for Progression Through T Cell Development.
Regulation of the Signal-Dependent E Protein HEBAlt Through a YYY Motif Is Required for Progression Through T Cell Development.
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DOI：
10.3389/fimmu.2022.848577
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发表时间：
2022
影响因子：
7.3
通讯作者：
Anderson, Michele K.
中科院分区：
文献类型：
--
作者：
Yoganathan, Kogulan;Yan, Anqi;Rocha, Juliana;Trotman-Grant, Ashton;Mohtashami, Mahmood;Wells, Lisa;Zuniga-Pfluecker, Juan Carlos;Anderson, Michele K.
E蛋白转录因子E2 A和HEB对许多发育过程至关重要，包括T细胞发育。我们已经表明，Tcf 12基因座产生了两种不同的HEB蛋白，具有交替（HEBAlt）和典型（HEBCan）N-末端结构域，它们在早期T细胞发育过程中共表达。虽然HEBCan的功能结构域已经被很好地研究，但HEBAlt特异性（Alt）结构域的性质一直不清楚。在这里，我们提供了令人信服的证据，Alt域提供了一个网站的细胞因子信号和E蛋白活性的分子整合。我们的研究结果表明，一个独特的YYY基序在Alt结构域的磷酸化增加HEBAlt活性的10倍，这种增加是依赖于Janus激酶活性。为了能够在T细胞环境中进行HEBAlt的体内研究，我们产生了ALT-Tg小鼠，其可以在Cre重组酶的存在下被诱导表达HA标记的HEBAlt编码盒。在Vav-iCre背景下对ALT-Tg小鼠进行的分析显示，ISP细胞与CD 8 + SP细胞的比率发生微小变化，脾脏中T细胞与B细胞的比率发生轻微变化，但胸腺、脾脏和骨髓淋巴细胞亚群在稳态下相当。然而，在0 P9-DL 4共培养物中T细胞发育的动力学分析揭示了当HEBAlt水平或活性增加时，早期T细胞发育的延迟和DN至DP转变的部分阻断。我们还观察到HEBCan和HEBAlt在蛋白质稳定性方面显示出显著差异，这些差异在胸腺细胞背景下得到解决。最后，蛋白质组学筛选确定STAT 1和Xpo 1是胸腺细胞中含HEBAlt复合物的潜在成员，这与JAK诱导的HEBAlt激活并伴有易位到细胞核一致。因此，我们的研究结果表明，Alt结构域赋予HEBAlt多个翻译后控制层，而HEBCan无法获得这些控制层，因此可以作为一个变阻器，在T细胞发育过程中，当细胞通过不同的胸腺信号环境时，调节E蛋白的活性水平。
The E protein transcription factors E2A and HEB are critical for many developmental processes, including T cell development. We have shown that the Tcf12 locus gives rise to two distinct HEB proteins, with alternative (HEBAlt) and canonical (HEBCan) N-terminal domains, which are co-expressed during early T cell development. While the functional domains of HEBCan have been well studied, the nature of the HEBAlt-specific (Alt) domain has been obscure. Here we provide compelling evidence that the Alt domain provides a site for the molecular integration of cytokine signaling and E protein activity. Our results indicate that phosphorylation of a unique YYY motif in the Alt domain increases HEBAlt activity by 10-fold, and that this increase is dependent on Janus kinase activity. To enable in vivo studies of HEBAlt in the T cell context, we generated ALT-Tg mice, which can be induced to express a HA-tagged HEBAlt coding cassette in the presence of Cre recombinases. Analysis of ALT-Tg mice on the Vav-iCre background revealed a minor change in the ratio of ISP cells to CD8+ SP cells, and a mild shift in the ratio of T cells to B cells in the spleen, but otherwise the thymus, spleen, and bone marrow lymphocyte subsets were comparable at steady state. However, kinetic analysis of T cell development in OP9-DL4 co-cultures revealed a delay in early T cell development and a partial block at the DN to DP transition when HEBAlt levels or activity were increased. We also observed that HEBCan and HEBAlt displayed significant differences in protein stability that were resolved in the thymocyte context. Finally, a proteomic screen identified STAT1 and Xpo1 as potential members of HEBAlt-containing complexes in thymocytes, consistent with JAK-induced activation of HEBAlt accompanied by translocation to the nucleus. Thus, our results show that the Alt domain confers access to multiple layers of post-translational control to HEBAlt that are not available to HEBCan, and thus may serve as a rheostat to tune E protein activity levels as cells move through different thymic signaling environments during T cell development.
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影响因子：
16.6
作者：
In TSH;Trotman-Grant A;Fahl S;Chen ELY;Zarin P;Moore AJ;Wiest DL;Zúñiga-Pflücker JC;Anderson MK
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