Promoter activity of the proliferating-cell nuclear antigen gene is associated with inducible CRE-binding proteins in interleukin 2-stimulated T lymphocytes

Danyang Huang, Pierette M. Shipman-Appasamy, Dana J. Orten, Steven H. Hinrichs, Michael B. Prystowsky

Research output: Contribution to journalArticle

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Abstract

The proliferating-cell nuclear antigen (PCNA) gene encodes an auxiliary factor of DNA polymerase delta and functions in DNA replication during S phase. It is expressed at much higher levels in proliferating cells than in quiescent cells. We have studied the regulatory role of the 5'-flanking sequence of the murine PCNA gene in interleukin 2 (IL-2)-responsive cloned T cells (L2). Analysis of a set of deletion constructs in transient transfection assays measuring heterologous reporter gene (luciferase) activity demonstrated that the 182-bp 5'-flanking region provides full promoter activity in IL-2-stimulated L2 cells. While many elements contribute to PCNA promoter strength in IL-2-stimulated cells, the largest decrease in activity occurred with deletion of the tandem CRE (cyclic AMP response element) binding sites located at nucleotides -37 to -52. With a gel mobility shift assay, several IL-2-inducible DNA-protein complexes were detected, including CREB (CRE-binding) and ATF1 (activating transcription factor) proteins that are specific for the PCNA-CRE sequence. Methylation interference analysis confirmed specific binding of these proteins to the CRE sites. Mutation at the PCNA-CRE motif abolishes IL-2-inducible binding and reduces substantially PCNA promoter activity. These results indicate that IL- 2-stimulated PCNA transcription may be partially mediated by these CRE- binding proteins.

Original languageEnglish (US)
Pages (from-to)4233-4243
Number of pages11
JournalMolecular and Cellular Biology
Volume14
Issue number6
StatePublished - Jun 1994

Fingerprint

Cyclic AMP Response Element-Binding Protein
Proliferating Cell Nuclear Antigen
Interleukin-2
T-Lymphocytes
Response Elements
Cyclic AMP
Genes
5' Flanking Region
Activating Transcription Factor 1
Activating Transcription Factors
DNA Polymerase III
Electrophoretic Mobility Shift Assay
Luciferases
DNA Replication
S Phase
Reporter Genes
Methylation
Transfection
Proteins
Nucleotides

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Promoter activity of the proliferating-cell nuclear antigen gene is associated with inducible CRE-binding proteins in interleukin 2-stimulated T lymphocytes. / Huang, Danyang; Shipman-Appasamy, Pierette M.; Orten, Dana J.; Hinrichs, Steven H.; Prystowsky, Michael B.

In: Molecular and Cellular Biology, Vol. 14, No. 6, 06.1994, p. 4233-4243.

Research output: Contribution to journalArticle

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