CCAAT/Enhancer-Binding Protein ε27 Antagonism of GATA-1 Transcriptional Activity in the Eosinophil Is Mediated by a Unique N-Terminal Repression Domain, Is Independent of Sumoylation and Does Not Require DNA Binding
Abstract
:1. Introduction
2. Results
2.1. C/EBPε and GATA-1 Bind In Vivo in Eosinophil Myelocytes to Their Functional Sites in the MBP1-P2 Promoter
2.2. Transduction with a TAT-C/EBPε27 Fusion Protein Inhibits GATA-1 Transactivation of the MBP1-P2 Promoter in CV-1 Cells and Expression of the Endogenous MBP1 Gene in AML14.3D10 Eosinophils
2.3. Repression Domains of C/EBPε27 Attenuate GATA-1 Transcriptional Activity: Both the Unique C/EBPε27 N-Terminus (RD27) and RDI Domains Inhibit GATA-1 Activity
2.4. C/EBPε Is Constitutively Sumoylated in Eosinophilic Myelocytes and in Heterologous Cells Transfected with a SUMO-1 Expression Vector
2.5. Over-Expression of SUMO-1 Has No Effect on C/EBPε27 Inhibition of GATA-1 Transactivation of the MBP1-P2 Promoter
2.6. Deletion of the DNA Binding Domain of C/EBPε27 Only Partially Relieves Repressor Activity for GATA-1
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Transfections
4.2. Plasmid Constructs
4.3. Chromatin Immunoprecipitation (ChIP)
4.4. Tat-C/EBPε Fusion Proteins and Cell Transductions
4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Stankiewicz, M.J.; Du, J.; Martinico, D.; Ackerman, S.J. CCAAT/Enhancer-Binding Protein ε27 Antagonism of GATA-1 Transcriptional Activity in the Eosinophil Is Mediated by a Unique N-Terminal Repression Domain, Is Independent of Sumoylation and Does Not Require DNA Binding. Int. J. Mol. Sci. 2021, 22, 12689. https://doi.org/10.3390/ijms222312689
Stankiewicz MJ, Du J, Martinico D, Ackerman SJ. CCAAT/Enhancer-Binding Protein ε27 Antagonism of GATA-1 Transcriptional Activity in the Eosinophil Is Mediated by a Unique N-Terminal Repression Domain, Is Independent of Sumoylation and Does Not Require DNA Binding. International Journal of Molecular Sciences. 2021; 22(23):12689. https://doi.org/10.3390/ijms222312689
Chicago/Turabian StyleStankiewicz, Monika J., Jian Du, Dominick Martinico, and Steven J. Ackerman. 2021. "CCAAT/Enhancer-Binding Protein ε27 Antagonism of GATA-1 Transcriptional Activity in the Eosinophil Is Mediated by a Unique N-Terminal Repression Domain, Is Independent of Sumoylation and Does Not Require DNA Binding" International Journal of Molecular Sciences 22, no. 23: 12689. https://doi.org/10.3390/ijms222312689