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Cyclic AMP/PKA‐triggered secondary transcrption networks associated with growth arrest and apoptosis in S49 lymphoma cells
Author(s) -
Wilderman Andrea C,
Insel Paul A,
Zambon Alex C
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.678.23
Subject(s) - creb , transcription factor , activator (genetics) , gene , electrophoretic mobility shift assay , microbiology and biotechnology , protein kinase a , biology , kinase , chemistry , biochemistry
The second messenger cyclic AMP (cAMP) acts via protein kinase A (PKA) to induce growth arrest and apoptosis in murine S49 lymphoma cells by mechanisms that are poorly understood. Here, we devised a bioinformatic method to predict the transcription factors (TFs) and their target genes that are activated or repressed downstream of cAMP/PKA. Our bioinformatic method identifies patterns of transcriptional co‐regulation by clustering and predicts the TFs that regulate the clusters by comparative genomic analysis of the 5′ regulatory regions for enrichment of TF binding sites. We used this methodology to analyze gene expression patterns of S49 cells treated for 0, 2, 6, and 24 hrs with the PKA‐activating analog 8‐CPT‐cAMP and found a potential role for a number of TFs and their target genes. We validated alterations of TF activity by electrophoretic mobility shift assay (EMSA). Increases in CREB activity reflect its role as a transcription activator of cAMP/PKA while increases in PPARγ, NFkB, and FOXO activity are associated with transcriptional down‐regulation. The findings indicate the utility of bioinformatic methods for predicting secondary transcriptional networks from gene expression data and define candidate TFs that regulate cAMP/PKA targets. (Supported by grants from NIH and the Lymphoma and Leukemia Society)