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Stimulation of NF-E2 DNA binding by CREB-binding protein (CBP)-mediated acetylation
1Division of Hematology, Children's Hospital of Philadelphia and the University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
The Journal of Biological Chemistry
|January 23, 2001
Summary
The cAMP-response element-binding protein (CREB)-binding protein (CBP) acetylates MafG, enhancing the DNA binding and transcriptional activity of the hematopoietic transcription factor NF-E2, crucial for blood cell development.
Area of Science:
- Molecular Biology
- Hematopoiesis
- Gene Regulation
Background:
- The transcription factor NF-E2 is vital for erythroid and megakaryocytic gene expression.
- CBP, a transcription cofactor, is known to mediate NF-E2 function.
Purpose of the Study:
- To investigate the specific role of CBP, a coactivator with acetyltransferase activity, in regulating NF-E2.
- To elucidate the mechanism by which CBP influences NF-E2 activity.
Main Methods:
- In vitro and in vivo interaction studies between NF-E2 subunits (p45, MafG) and CBP.
- Acetylation assays to determine CBP's effect on MafG and p45.
- Immunoprecipitation with anti-acetyl lysine antibodies to detect in vivo MafG acetylation.
- Transfection experiments to assess CBP's role in MafG acetylation and NF-E2 activity.
Main Results:
- Both p45 and MafG subunits of NF-E2 interact with CBP.
- CBP specifically acetylates MafG, not p45, primarily in MafG's basic region.
- MafG is acetylated in vivo in erythroid cells, and CBP enhances this acetylation.
- MafG acetylation by CBP significantly increases NF-E2 DNA binding and transcriptional activation.
Conclusions:
- CBP-mediated acetylation of MafG is a key mechanism for regulating NF-E2 activity.
- NF-E2 likely recruits CBP to erythroid/megakaryocytic promoters for transcriptional regulation.
- Regulation involves both chromatin modification and direct modulation of transcription factor activity.