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Updated: Aug 19, 2026
Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis
Published on: April 1, 2022
Identification of novel CBP interacting proteins in embryonic orofacial tissue
Xiaolong Yin1, Dennis R Warner, Emily A Roberts
1Department of Molecular, Cellular and Craniofacial Biology, University of Louisville Birth Defects Center, ULSD Louisville, KY 40292, USA.
Abstract:
cAMP response element-binding protein (CREB)-binding protein (CBP) plays an important role as a general co-integrator of multiple signaling pathways and interacts with a large number of transcription factors and co-factors, through its numerous protein-binding domains. To identify nuclear factors associated with CBP in developing orofacial tissue, a yeast two-hybrid screen of a cDNA library derived from orofacial tissue from gestational day 11 to 13 mouse embryos was conducted. Using the carboxy terminus (amino acid residues 1676-2441) of CBP as bait, several novel proteins that bind CBP were identified, including an Msx-interacting-zinc finger protein, CDC42 interaction protein 4/thyroid hormone receptor interactor 10, SH3-domain GRB2-like 1, CCR4-NOT transcription complex subunit 3, adaptor protein complex AP-1 beta1 subunit, eukaryotic translation initiation factor 2B subunit 1 (alpha), and cyclin G-associated kinase. Results of the yeast two-hybrid screen were confirmed by glutathione S-transferase pull-down assays. The identification of these proteins as novel CBP-binding partners allows exploration of new mechanisms by which CBP regulates and integrates diverse cell signaling pathways.
Insights
Researchers identified novel nuclear proteins interacting with cAMP response element-binding protein (CREB)-binding protein (CBP) in developing orofacial tissue. This discovery offers new insights into CBP
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- cAMP response element-binding protein (CREB)-binding protein (CBP) is a crucial co-integrator of signaling pathways.
- CBP interacts with numerous transcription factors and co-factors via its protein-binding domains.
Purpose of the Study:
- To identify nuclear factors that associate with CBP in developing orofacial tissue.
- To explore novel CBP-binding partners involved in orofacial development.
Main Methods:
- Yeast two-hybrid screen using the carboxy terminus of CBP as bait.
- Utilized a cDNA library from gestational day 11-13 mouse embryonic orofacial tissue.
- Confirmed interactions using glutathione S-transferase pull-down assays.
Main Results:
- Identified several novel CBP-binding proteins, including Msx-interacting-zinc finger protein and eukaryotic translation initiation factor 2B subunit 1 (alpha).
- Other identified proteins include CDC42 interaction protein 4/thyroid hormone receptor interactor 10, SH3-domain GRB2-like 1, CCR4-NOT transcription complex subunit 3, AP-1 beta1 subunit, and cyclin G-associated kinase.
- Validated findings through glutathione S-transferase pull-down assays.
Conclusions:
- This study identifies novel CBP-binding partners in orofacial tissue.
- These findings open new avenues for understanding how CBP regulates diverse cell signaling pathways during development.
