Identification of an essential cis-element near the transcription start site for transcriptional activation of the proliferating cell nuclear antigen gene.
Interleukin 2 (IL-2) stimulates T lymphocyte proliferation and induces the expression of proliferating cell nuclear antigen (PCNA), a processivity factor for DNA polymerase delta. Previously, deletion analysis suggested cis-element(s) in the proximal region of the PCNA promoter (-40 to +143) are required for IL-2 induction in cloned T lymphocytes. The sequence 5'-TTGCGGGC-3' located at +10 to +17 is similar to the E2F consensus binding site and is required for optimal PCNA promoter activity. In IL-2-stimulated T cells, nuclear proteins are induced to bind to this sequence as demonstrated using electrophoretic mobility shift assay (EMSA), competition EMSA, and methylation interference analysis. A 180-kDa polypeptide was detected by UV cross-linking to bind specifically to the PCNA E2F-like sequence. Our data indicate that the protein bound to the PCNA E2F-like site is not one of the transcription factor E2F proteins. Our results demonstrate that the E2F-like sequence and the protein(s) binding to it are required for optimal PCNA promoter activity and IL-2 induction of PCNA expression.
Huang
D Y
DY
Graduate group of Molecular Biology, University of Pennsylvania, School of Medicine, Philadelphia 19104, USA.
Prystowsky
M B
MB
D000818:Animals; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D007376:Interleukin-2 / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D017421:Sequence Analysis; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D015533:Transcriptional Activation
D007376:Interleukin-2; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen