Morris G F, Mathews M B
Cold Spring Harbor Laboratory, New York 11724.
J Biol Chem. 1990 Sep 25;265(27):16116-25.
The levels of the mRNA for the proliferating cell nuclear antigen (PCNA), a DNA replication factor, increase upon growth stimulation of quiescent cells. To study the transcriptional aspect of this response, we have cloned a PCNA gene fragment from size-fractionated human placental DNA. This fragment contains 1269 nucleotides upstream from the PCNA transcriptional start site and includes an Alu sequence that is transcribed in vitro. The PCNA genomic DNA promotes transcription of a linked heterologous reporter gene in HeLa and 293 cells. Transient expression assays and in vitro transcription analyses showed that 249 nucleotides of upstream sequence are sufficient for full promoter activity in HeLa cells, whereas only 172 nucleotides are needed in 293 cells. Co-transfection with a plasmid expressing the adenovirus E1 gene transactivates the PCNA promoter in HeLa cells. An E1-responsive element maps in the 85-nucleotide region immediately upstream of the site of transcription initiation.
增殖细胞核抗原（PCNA）是一种DNA复制因子，静止细胞在生长刺激后，其mRNA水平会升高。为了研究这种反应的转录方面，我们从大小分级的人胎盘DNA中克隆了一个PCNA基因片段。该片段包含PCNA转录起始位点上游1269个核苷酸，并包含一个在体外可转录的Alu序列。PCNA基因组DNA可促进HeLa和293细胞中相连的异源报告基因的转录。瞬时表达分析和体外转录分析表明，249个核苷酸的上游序列足以在HeLa细胞中实现完整的启动子活性，而在293细胞中仅需要172个核苷酸。与表达腺病毒E1基因的质粒共转染可激活HeLa细胞中的PCNA启动子。一个E1反应元件位于转录起始位点上游紧邻的85个核苷酸区域内。
