UVB RADIATION-ACTIVATED GENES INDUCED BY TRANSCRIPTIONAL AND POSTTRANSCRIPTIONAL MECHANISMS IN RAT KERATINOCYTES
UVB RADIATION-ACTIVATED GENES INDUCED BY TRANSCRIPTIONAL AND POSTTRANSCRIPTIONAL MECHANISMS IN RAT KERATINOCYTES
复制标题
DOI：
10.1152/ajpcell.1995.268.4.c846
复制
发表时间：
1995-04-01
影响因子：
5.5
通讯作者：
DRUCKER, DJ
中科院分区：
文献类型：
--
作者：
ROSEN, CF;POON, R;DRUCKER, DJ
紫外线 B (UVB) 辐射在基因表达调节中所利用的机制以及 WE 信号传输的遗传靶标仍不完全清楚。为了阐明哺乳动物细胞中 WE 激活的机制和靶标，我们使用差异消减的富含 UVB 的 cDNA 探针筛选了角质形成细胞 cDNA 文库。分离出 23 个 UVB 诱导的 cDNA 克隆，包括角蛋白、延伸因子 1 α、铁蛋白重链、硫醇转移酶、细胞周期蛋白 G、cornifin、cellubrevin、poly(A) 结合蛋白和 surfeit 基因座的 cDNA。 UVB 暴露后最大 RNA 诱导的时间动力学是异质的，在 WB 辐射后 1 到 24 小时之间变化。对基因表达调控的分析表明，大多数 UVB 诱导的 mRNA 水平也由血清和放线菌酮独立诱导，这是先前描述的 DNA 损伤诱导基因和直接早期基因家族成员的特征。与立即早期基因研究的结果相反，用血清和放线菌酮处理角质形成细胞仅导致一种 mRNA 转录物的超诱导。核连续分析表明，UVB 辐射增加了 23 个基因中 8 个基因的转录率，表明 UVB 辐射利用转录和转录后机制来调节角质形成细胞基因表达。一组 UVB 诱导的角质形成细胞基因的鉴定对于表征对 WE 辐射的基因组反应以及分析 UVB 基因表达调节的分子机制应该是有用的。
The mechanisms utilized by ultraviolet B (UVB) radiation in the regulation of gene expression, as well as the genetic targets for transmission of the WE signal, remain incompletely understood. To elucidate the mechanisms and targets for WE activation in mammalian cells, we screened a keratinocyte cDNA library with differentially subtracted UVB-enriched cDNA probes. Twenty-three UVB-induced cDNA clones were isolated, including cDNAs for keratin, elongation factor-1 alpha, ferritin heavy chain, thioltransferase, cyclin G, cornifin, cellubrevin, poly(A) binding protein, and the surfeit locus. The temporal kinetics of maximal RNA induction following UVB exposure were heterogeneous, varying from 1 to 24 h post-WB radiation. Analysis of the regulation of gene expression demonstrated that the levels of most UVB-induced mRNAs were also independently induced by serum and cycloheximide, features previously described for genes induced by DNA damage and members of the immediate early gene family. In contrast to results from studies of immediate early genes, treatment of keratinocytes with both serum and cycloheximide resulted in superinduction of only one mRNA transcript. Nuclear run-on assays demonstrated that UVB radiation increased the transcription rate in 8 of 23 genes, suggesting that UVB radiation utilizes both transcriptional and posttranscriptional mechanisms for the modulation of keratinocyte gene expression. The identification of a group of UVB-inducible keratinocyte genes should prove useful for the characterization of the genomic response to WE radiation and the analysis of the molecular mechanisms underlying the UVB regulation of gene expression.