Harrigan M T, Baughman G, Campbell N F, Bourgeois S
Regulatory Biology Laboratory, Salk Institute for Biological Studies, San Diego, California 92138.
Mol Cell Biol. 1989 Aug;9(8):3438-46. doi: 10.1128/mcb.9.8.3438-3446.1989.
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 5 h 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.
糖皮质激素和环磷酸腺苷(cAMP)通过未知机制对小鼠T淋巴细胞的增殖和活力产生显著影响。为了鉴定可能参与淋巴细胞中糖皮质激素诱导反应的基因产物,我们构建了一个λ cDNA文库,该文库由用糖皮质激素和福斯可林处理5小时的小鼠胸腺瘤WEHI-7TG细胞制备而成。用富集了两种药物诱导序列的扣除cDNA探针筛选该文库,分离出代表11个不同诱导基因的cDNA克隆。对每个cDNA克隆检测其在BALB/c小鼠组织中的表达模式。我们鉴定出两个与仅在胸腺中检测到的mRNA杂交的克隆。还鉴定出其他克隆,它们在心脏、脑、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。检测了每种基因受地塞米松和福斯可林诱导的动力学。大多数cDNA克隆与受糖皮质激素和福斯可林调节的mRNA杂交,两个仅受糖皮质激素调节,三个与需要两种药物诱导的mRNA杂交。用环己酰亚胺抑制蛋白质合成导致所有可被糖皮质激素诱导的mRNA的诱导。对11个cDNA中的4个进行的初步序列分析表明,两个cDNA代表以前未描述的基因,而另外两个分别对应于小鼠VL30逆转录病毒样元件和硫酸软骨素蛋白聚糖核心蛋白的小鼠同源物。