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Published on: May 1, 2013
Use of an array technology for profiling and comparing transcription factors activated by TNFalpha and PMA in HeLa
Xin Jiang1, Michael Norman, Xianqiang Li
1Panomics, Inc., 2003 East Bayshore Road, Redwood City, CA 94063, USA. xjiang@panomics.com
Abstract:
Multiple signal transduction pathways are generally triggered simultaneously by a single extracellular stimulus. As a result, multiple transcription factors (TFs) can be activated downstream to mediate the inducible expression of target genes. Profiling the activation of all TFs will aid in the dissection of the numerous pathways of signal transduction. Tumor necrosis factor alpha (TNFalpha) and phorbol 12-myristate 13-acetate (PMA) mediate many biological functions, including cell proliferation and apoptosis, by stimulating signaling pathways. Two TFs, nuclear factor kappaB (NFkappaB) and activating factor 1 (AP1), have been identified as targets of both TNFalpha and PMA activation. Here, we describe the use of a protein/DNA array system to identify additional TFs activated by TNFalpha and PMA in HeLa cells. From a total of 150 targeted TFs, six-CREB, E2F, CETP/CRE, c-Rel, MSP1, and Pax6-were identified whose activities, like NFkappaB and AP1, were regulated by both TNFalpha- and PMA-induced pathways. Interestingly, the TF E47 was shown to be specifically activated by TNFalpha but was not affected by treatment with PMA. In addition, GATA, NF-E1, and ISRE were shown to be specifically activated by PMA but not TNFalpha. These findings suggest that TNFalpha and PMA both stimulate unique signaling pathways while mediating transcriptional activation through common pathways.
Insights
This study profiles transcription factor (TF) activation by tumor necrosis factor alpha (TNFα) and phorbol 12-myristate 13-acetate (PMA). A protein/DNA array identified common and unique TFs activated by these stimuli, revealing insights into signal transduction pathways.
Area of Science:
- Molecular Biology
- Cell Signaling
- Genomics
Background:
- Extracellular stimuli activate multiple signal transduction pathways simultaneously.
- Transcription factors (TFs) mediate inducible gene expression downstream of these pathways.
- Understanding TF activation is crucial for dissecting complex signaling networks.
Purpose of the Study:
- To identify transcription factors (TFs) activated by tumor necrosis factor alpha (TNFα) and phorbol 12-myristate 13-acetate (PMA) in HeLa cells.
- To profile the activation of a broad range of TFs using a protein/DNA array system.
- To differentiate common and unique TF activation patterns induced by TNFα and PMA.
Main Methods:
- Utilized a protein/DNA array system to screen 150 targeted transcription factors (TFs).
- Treated HeLa cells with TNFα and PMA to stimulate signaling pathways.
- Analyzed TF activation profiles in response to each stimulus.
Main Results:
- Identified six TFs (CREB, E2F, CETP/CRE, c-Rel, MSP1, and Pax6) activated by both TNFα and PMA, similar to NFκB and AP1.
- Discovered E47 was specifically activated by TNFα but not PMA.
- Found GATA, NF-E1, and ISRE were specifically activated by PMA but not TNFα.
Conclusions:
- TNFα and PMA activate distinct signaling pathways leading to transcriptional regulation.
- Both stimuli converge on common pathways to regulate specific TFs.
- The study provides a comprehensive profile of TF activation by TNFα and PMA, highlighting pathway specificity and convergence.
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