Yeast one-hybrid screen of a thymus epithelial library identifies ZBTB7A as a regulator of thymic insulin expression.
Yeast one-hybrid screen of a thymus epithelial library identifies ZBTB7A as a regulator of thymic insulin expression.
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胸腺上皮文库的酵母单杂交筛选将 ZBTB7A 鉴定为胸腺胰岛素表达的调节剂。
DOI：
10.1016/j.molimm.2013.05.238
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发表时间：
2013
影响因子：
3.6
通讯作者：
Polychronakos,Constantin
中科院分区：
文献类型：
--
作者：
St-Jean,JulienR;Ounissi-Benkalha,Houria;Polychronakos,Constantin
胰岛素自身耐受性在很大程度上是由胸腺髓质上皮细胞（mTECs）表达少量胰岛素来保证的。胸腺胰岛素表达的调节不同于胰腺，其治疗操作在预防1型糖尿病（T1 D）中可能发挥重要作用。mTEC核环境中涉及的转录调节因子的知识对于开发此类疗法是必不可少的。使用酵母单杂交（Y1 H）方法以鉴定这样的mTEC特异性核蛋白。我们使用了一个目标组成的人胰岛素基因启动子连接到上游III类VNTR等位基因，这是与保护T1 D和更高的胸腺胰岛素表达，和cDNA文库从我们的胰岛素生产小鼠mTEC线。Y1 H筛选允许鉴定11种蛋白质。体外分析用于确认和定量与单独的人胰岛素基因启动子或与I类或III类VNTR等位基因连接的蛋白质-DNA结合，并鉴定转录因子ZBTB 7A、JUN和EWSR 1为强相互作用伴侣。所有三种蛋白质都可以诱导转染的HEK-293细胞中的胰岛素表达，但ZBTB 7A提供了最稳健的结果，特别是在AIRE存在下，来自III类VNTR等位基因驱动的共转染报告基因的胰岛素mRNA水平额外增加11倍。因此，ZBTB 7A被鉴定为调节胸腺胰岛素表达的强有力的候选者。
Insulin self-tolerance is, to a large extent, assured by the expression of small quantities of insulin by medullary thymic epithelial cells (mTECs). Regulation of thymic insulin expression differs from that in pancreas and its therapeutic manipulation could play an important role in the prevention of type 1 diabetes (T1D). Knowledge of the transcriptional regulators involved in the mTEC nuclear environment is essential for the development of such therapeutics. The yeast one-hybrid (Y1H) approach was used in order to identify such mTEC-specific nuclear proteins. We used a target composed of the human insulin gene promoter joined to the upstream class III VNTR allele, which is associated with both protection from T1D and higher thymic insulin expression, and a cDNA library from our insulin-producing mouse mTEC line. The Y1H screening allowed the identification of eleven proteins. Anin vitroassay was used to confirm and quantify protein–DNA binding to the human insulin gene promoter alone or joined to a class I or class III VNTR allele, and identified the transcription factors ZBTB7A, JUN and EWSR1 as strong interacting partners. All three proteins could induce insulin expression in transfected HEK-293 cells, but ZBTB7A provided the most robust results especially in the presence of AIRE, with an additional 11-fold increase of the insulin mRNA levels from a co-transfected reporter driven by the class III VNTR allele. Thus, ZBTB7A is identified as a strong candidate for regulation of thymic insulin expression.
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影响因子：
64.8
作者：
Kent, SC;Chen, YH;Hafler, DA
通讯作者：
Hafler, DA
DOI：
10.1093/bfgp/ell015
发表时间：
2006-03-01
期刊：
Briefings in Functional Genomics & Proteomics
影响因子：
--
作者：
Law, Warren J.;Cann, Kendra L.;Hicks, Geoffrey G.
通讯作者：
Hicks, Geoffrey G.
影响因子：
64.8
作者：
Nakayama, M;Abiru, N;Eisenbarth, GS
通讯作者：
Eisenbarth, GS
影响因子：
11.4
作者：
J. Tong;E. Duprez;M. Lanotte
通讯作者：
J. Tong;E. Duprez;M. Lanotte
影响因子：
8.2
作者：
Levi, D.;Polychronakos, C.
通讯作者：
Polychronakos, C.