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Promoter activity of the proliferating-cell nuclear antigen gene is associated with inducible CRE-binding proteins in interleukin 2-stimulated T lymphocytes.
Huang, D; Shipman-Appasamy, P M; Orten, D J; Hinrichs, S H; Prystowsky, M B.
Afiliação
  • Huang D; Department of Pathology, School of Medicine, University of Pennsylvania, Philadelphia 19104.
Mol Cell Biol ; 14(6): 4233-43, 1994 Jun.
Article em En | MEDLINE | ID: mdl-7910946
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Nucleares / Linfócitos T / Interleucina-2 / Regiões Promotoras Genéticas / Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico Tipo de estudo: Risk_factors_studies Limite: Animals Idioma: En Ano de publicação: 1994 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Nucleares / Linfócitos T / Interleucina-2 / Regiões Promotoras Genéticas / Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico Tipo de estudo: Risk_factors_studies Limite: Animals Idioma: En Ano de publicação: 1994 Tipo de documento: Article