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Identification of a multiprotein complex interacting with the c-fos serum response element
I de Belle1, P R Walker, I C Smith
1Cellular Oncology Group, Institute for Biological Sciences, National Research Council, Ottawa, Ontario, Canada.
Molecular and Cellular Biology
|May 1, 1991
Summary
Researchers identified novel proteins interacting with the serum response element (SRE) in the c-fos gene. Phosphorylation governs the formation of a multiprotein complex essential for c-fos gene transcriptional activation.
Area of Science:
- Molecular Biology
- Gene Regulation
- Biochemistry
Background:
- The serum response element (SRE) is crucial for stimulating the c-fos gene by serum and growth factors.
- Understanding the proteins that bind to the SRE is key to deciphering c-fos gene regulation.
Purpose of the Study:
- To identify and characterize nuclear proteins that bind to the SRE sequence of the c-fos gene.
- To investigate the role of protein phosphorylation in the formation of SRE-protein complexes.
Main Methods:
- Mobility shift assay
- Southwestern (DNA-protein) blotting
- Sequence-specific affinity chromatography using a labeled SRE oligonucleotide probe.
Main Results:
- Identified nuclear proteins of 36, 45, 62, 67, 72, and 112 kDa that interact with the SRE.
- 36, 67, and 112 kDa proteins exhibit DNA-binding properties; others may interact via protein-protein interactions.
- Phosphorylation is critical for the formation of the SRE-protein complex, impacting transcriptional activation of c-fos.
- Discovered novel proteins (36, 45, 72, 112 kDa) involved in c-fos gene regulation.
Conclusions:
- Transcriptional activation of the c-fos gene at the SRE requires a multiprotein complex.
- The formation of this complex is regulated by protein phosphorylation.
- Novel protein factors contribute to the complex regulation of c-fos gene expression in vivo.