Tags

Type your tag names separated by a space and hit enter

Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site.
Cell Rep. 2019 01 22; 26(4):845-854.e6.CR

Abstract

Notch is a conserved signaling pathway that is essential for metazoan development and homeostasis; dysregulated signaling underlies the pathophysiology of numerous human diseases. Receptor-ligand interactions result in gene expression changes, which are regulated by the transcription factor RBPJ. RBPJ forms a complex with the intracellular domain of the Notch receptor and the coactivator Mastermind to activate transcription, but it can also function as a repressor by interacting with corepressor proteins. Here, we determine the structure of RBPJ bound to the corepressor SHARP and DNA, revealing its mode of binding to RBPJ. We tested structure-based mutants in biophysical and biochemical-cellular assays to characterize the role of RBPJ as a repressor, clearly demonstrating that RBPJ mutants deficient for SHARP binding are incapable of repressing transcription of genes responsive to Notch signaling in cells. Altogether, our structure-function studies provide significant insights into the repressor function of RBPJ.

Authors+Show Affiliations

Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.Institute of Biochemistry, University of Giessen, Giessen, Germany.Department of Internal Medicine I, Center for Internal Medicine, University Medical Center Ulm, 89081 Ulm, Germany.Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.Institute of Biochemistry, University of Giessen, Giessen, Germany.Institute of Biochemistry, University of Giessen, Giessen, Germany.Department of Internal Medicine I, Center for Internal Medicine, University Medical Center Ulm, 89081 Ulm, Germany.Institute of Biochemistry, University of Giessen, Giessen, Germany.Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH, USA. Electronic address: kovallra@ucmail.uc.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

30673607

Citation

Yuan, Zhenyu, et al. "Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site." Cell Reports, vol. 26, no. 4, 2019, pp. 845-854.e6.
Yuan Z, VanderWielen BD, Giaimo BD, et al. Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site. Cell Rep. 2019;26(4):845-854.e6.
Yuan, Z., VanderWielen, B. D., Giaimo, B. D., Pan, L., Collins, C. E., Turkiewicz, A., Hein, K., Oswald, F., Borggrefe, T., & Kovall, R. A. (2019). Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site. Cell Reports, 26(4), 845-e6. https://doi.org/10.1016/j.celrep.2018.12.097
Yuan Z, et al. Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site. Cell Rep. 2019 01 22;26(4):845-854.e6. PubMed PMID: 30673607.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site. AU - Yuan,Zhenyu, AU - VanderWielen,Bradley D, AU - Giaimo,Benedetto Daniele, AU - Pan,Leiling, AU - Collins,Courtney E, AU - Turkiewicz,Aleksandra, AU - Hein,Kerstin, AU - Oswald,Franz, AU - Borggrefe,Tilman, AU - Kovall,Rhett A, PY - 2018/05/31/received PY - 2018/11/05/revised PY - 2018/12/21/accepted PY - 2019/1/24/entrez PY - 2019/1/24/pubmed PY - 2020/3/7/medline KW - CSL KW - MINT KW - Notch signaling KW - RBPJ KW - SHARP KW - SPEN KW - X-ray crystallography KW - isothermal titration calorimetry KW - signal transduction KW - transcriptional regulation SP - 845 EP - 854.e6 JF - Cell reports JO - Cell Rep VL - 26 IS - 4 N2 - Notch is a conserved signaling pathway that is essential for metazoan development and homeostasis; dysregulated signaling underlies the pathophysiology of numerous human diseases. Receptor-ligand interactions result in gene expression changes, which are regulated by the transcription factor RBPJ. RBPJ forms a complex with the intracellular domain of the Notch receptor and the coactivator Mastermind to activate transcription, but it can also function as a repressor by interacting with corepressor proteins. Here, we determine the structure of RBPJ bound to the corepressor SHARP and DNA, revealing its mode of binding to RBPJ. We tested structure-based mutants in biophysical and biochemical-cellular assays to characterize the role of RBPJ as a repressor, clearly demonstrating that RBPJ mutants deficient for SHARP binding are incapable of repressing transcription of genes responsive to Notch signaling in cells. Altogether, our structure-function studies provide significant insights into the repressor function of RBPJ. SN - 2211-1247 UR - https://www.unboundmedicine.com/medline/citation/30673607/Structural_and_Functional_Studies_of_the_RBPJ_SHARP_Complex_Reveal_a_Conserved_Corepressor_Binding_Site_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S2211-1247(18)32057-6 DB - PRIME DP - Unbound Medicine ER -