Analysis of the proliferating cell nuclear antigen promoter and its response to adenovirus early region 1.
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.
Morris
G F
GF
Cold Spring Harbor Laboratory, New York 11724.
Mathews
M B
MB
D017456:Adenovirus Early Proteins; D000260:Adenoviruses, Human / Q000235:genetics; D000951:Antigens, Neoplasm / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D004261:DNA Replication; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D015345:Oligonucleotide Probes; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
D017456:Adenovirus Early Proteins; D000951:Antigens, Neoplasm; D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D009856:Oncogene Proteins, Viral; D018809:Proliferating Cell Nuclear Antigen; D015500:Chloramphenicol O-Acetyltransferase