ISOLATION AND CHARACTERIZATION OF GLUCOCORTICOID-INDUCED AND CYCLIC AMP-INDUCED GENES IN T-LYMPHOCYTES
ISOLATION AND CHARACTERIZATION OF GLUCOCORTICOID-INDUCED AND CYCLIC AMP-INDUCED GENES IN T-LYMPHOCYTES
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DOI：
10.1128/mcb.9.8.3438
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发表时间：
1989-08-01
影响因子：
5.3
通讯作者：
BOURGEOIS, S
中科院分区：
文献类型：
--
作者：
HARRIGAN, MT;BAUGHMAN, G;BOURGEOIS, S
糖皮质激素和环AMP通过未知的机制对小鼠T淋巴细胞的增殖和活力产生显著影响。为了鉴定可能参与糖皮质激素诱导淋巴细胞反应的基因产物，我们构建了一个。lambda。小鼠胸腺瘤WEHI-7TG细胞经糖皮质激素和福斯可林处理5h制备cDNA文库。用富集两种药物诱导序列的减法cDNA探针对文库进行筛选，分离出11个不同诱导基因的cDNA克隆。检测每个cDNA克隆在BALB/c小鼠组织中的表达模式。我们已经鉴定了两个克隆，它们与胸腺中检测到的mrna杂交。其他克隆在心脏、大脑、大脑和胸腺或淋巴组织（脾脏和胸腺）中表现出组织特异性基因表达。检测了地塞米松和福斯克林对各基因的诱导动力学。大多数cDNA克隆杂交到受糖皮质激素和forskolin调节的mrna，两个仅受糖皮质激素调节，三个杂交到需要两种药物诱导的mrna。环己亚胺对蛋白质合成的抑制导致糖皮质激素诱导的所有mrna的诱导。对11个cdna中的4个的初步序列分析表明，两个cdna代表先前未描述的基因，而另外两个对应于小鼠VL30逆转录病毒样元件和小鼠硫酸软骨素蛋白多糖核心蛋白的同源物。
Glucocorticoids and cyclic AMP exert dramatic effects on the proliferation and viability of murine T lymphocytes through unknown mechanisms. To identify gene products which might be involved in glucocorticoid-induced responses in lymphoid cells, we constructed a .lambda.cDNA library prepared from murine thymoma WEHI-7TG cells treated for 5h with glucocorticoids and forskolin. The library was screened with a subtracted cDNA probe enriched for sequences induced by the two drugs, and cDNA clones representing 11 different inducible genes were isolated. The pattern of expression in BALB/c mouse tissues was examined for each cDNA clone. We have identified two clones that hybridized to mRNAs detected exclusively in the thymus. Other clones were identified that demonstrated tissue-specific gene expression in heart, brain, brain and thymus, or lymphoid tissue (spleen and thymus). The kinetics of induction by dexamethasone and forskolin were examined for each gene. The majority of the cDNA clones hybridized to mRNAs that were regulated by glucocorticoids and forskolin, two were regulated only by glucocorticoids, and three hybridized to mRNAs that required both drugs for induction. Inhibition of protein synthesis by cycloheximide resulted in the induction of all mRNAs that were inducible by glucocorticoids. Preliminary sequence analysis of four of the 11 cDNAs suggests that two cDNAs represent previously undescribed genes while two others correspond to the mouse VL30 retrovirus-like element and the mouse homolog of chondroitin sulfate proteoglycan core protein.