Transcriptional and posttranscriptional regulation of the proliferating cell nuclear antigen gene.
Transcriptional and posttranscriptional regulation of the proliferating cell nuclear antigen gene.
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增殖细胞核抗原基因的转录和转录后调控。
DOI：
10.1128/mcb.10.7.3289-3296.1990
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发表时间：
1990
影响因子：
5.3
通讯作者：
Baserga,R
中科院分区：
文献类型：
--
作者：
Chang,CD;Ottavio,L;Travali,S;Lipson,KE;Baserga,R
增殖细胞核抗原 (PCNA) 基因的稳态 mRNA 水平受到生长调节。在之前的一篇论文（L. Ottavio, C.-D. Chang, M. G. Rizzo, S. Travali, C. Casadevall, and R. Baserga, Mol. Cell. Biol. 10:303-309, 1990）中，我们报道了内含子（尤其是内含子 4）参与 PCNA 基因的生长调节。我们现在已经研究了稳定转染至 BALB/c 3T3 细胞中的人 PCNA 基因 5' 侧翼序列的作用。测试了不同长度的启动子（从帽位点上游的 –2856 到 –45）。除 AatII 启动子 (–45) 之外的所有启动子，包括短 HpaII 启动子 (–210)，都足以对血清、血小板衍生生长因子以及较小程度的表皮生长因子产生反应。没有构建体对胰岛素或贫血小板血浆有反应。 AatII启动子几乎没有可检测到的活性。还通过两种方法测定了携带各种人类 PCNA 基因构建体的 BALB/c 3T3 细胞中的转录活性：连续转录和逆转录聚合酶链式反应（后者测量异源核 RNA [hnRNA] 稳态水平）。尽管刺激后hnRNA的水平要高得多，但G0细胞和血清刺激的细胞之间PCNA基因的转录率差异很小。在携带没有内含子4和5的人PCNA基因的G0细胞中，转录率和hnRNA水平都很高。结合 mRNA 半衰期的数据，这些结果表明血清刺激后 PCNA mRNA 水平的调节存在转录后成分，但转录调节是由内含子 4 进行的。
The steady-state mRNA levels of the proliferating cell nuclear antigen (PCNA) gene are growth regulated. In a previous paper (L. Ottavio, C.-D. Chang, M. G. Rizzo, S. Travali, C. Casadevall, and R. Baserga, Mol. Cell. Biol. 10:303-309, 1990), we reported that introns (especially intron 4) participate in growth regulation of the PCNA gene. We have now investigated the role of the 5'-flanking sequence of the human PCNA gene stably transfected into BALB/c 3T3 cells. Promoters of different lengths (from –2856 to –45 upstream of the cap site) were tested. All promoters except theAatII promoter (–45), including a shortHpaII promoter (–210), were sufficient for a response to serum, platelet-derived growth factor, and to a lesser extent epidermal growth factor. No construct responded to insulin or platelet-poor plasma. TheAatII promoter had little detectable activity. Transcriptional activity was also determined in BALB/c 3T3 cells carrying various constructs of the human PCNA gene by two methods: run-on transcription and reverse transcription-polymerase chain reaction (the latter measuring the heterogeneous nuclear RNA [hnRNA] steady-state levels). There was very little difference in the rate of transcription of the PCNA gene between G0cells and serum-stimulated cells, although the levels of hnRNA were much higher after stimulation. In G0cells carrying a human PCNA gene without introns 4 and 5, both transcription rate and hnRNA levels were high. Together with data on the mRNA half-life, these results suggest a posttranscriptional component in the regulation of PCNA mRNA levels after serum stimulation but a transcriptional regulation by intron 4.