Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site

Yuan, Zhenyu; VanderWielen, Bradley D.; Giaimo, Benedetto Daniele; Pan, Leiling; Collins, Courtney E.; Turkiewicz, Aleksandra; Hein, Kerstin; Oswald, Franz; Borggrefe, Tilman; Kovall, Rhett A.

doi:10.1016/j.celrep.2018.12.097

Title: Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site

Journal Article · Tue Jan 22 00:00:00 EST 2019 · Cell Reports

DOI:https://doi.org/10.1016/j.celrep.2018.12.097· OSTI ID:1569868

Yuan, Zhenyu ^[1]; VanderWielen, Bradley D. ^[1]; Giaimo, Benedetto Daniele ^[2]; Pan, Leiling ^[3]; Collins, Courtney E. ^[1]; Turkiewicz, Aleksandra ^[2]; Hein, Kerstin ^[2];

^[3];

^[2]; Kovall, Rhett A. ^[1]

Univ. of Cincinnati, OH (United States)
Univ. of Giessen (Germany)
University Medical Center, Ulm (Germany)

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.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC); National Institutes of Health (NIH); German Research Foundation (DFG); Excellence Cluster for Cardio Pulmonary System (ECCPS); University Medical Center Giessen and Marburg (UKGM); Federal Ministry of Education and Research, Research Nucleus SyStAR

Grant/Contract Number:: AC02-06CH11357; 5R01CA178974; TRR81; BO 1639/9-1; SFB1074/A3; GRK 2253 – HEIST

OSTI ID:: 1569868

Journal Information:: Cell Reports, Vol. 26, Issue 4; ISSN 2211-1247

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

Citation Metrics:

Cited by: 28 works

Citation information provided by
Web of Science

References (47)

Structural and functional analysis of the repressor complex in the Notch signaling pathway of Drosophila melanogaster Maier, Dieter; Kurth, Patricia; Schulz, Adriana Molecular Biology of the Cell, Vol. 22, Issue 17 https://doi.org/10.1091/mbc.e11-05-0420	journal	September 2011
Structure and Function of the CSL-KyoT2 Corepressor Complex: A Negative Regulator of Notch Signaling Collins, Kelly J.; Yuan, Zhenyu; Kovall, Rhett A. Structure, Vol. 22, Issue 1 https://doi.org/10.1016/j.str.2013.10.010	journal	January 2014
Histone variant H2A.Z deposition and acetylation directs the canonical Notch signaling response Giaimo, Benedetto Daniele; Ferrante, Francesca; Vallejo, Diana M. Nucleic Acids Research, Vol. 46, Issue 16 https://doi.org/10.1093/nar/gky551	journal	July 2018
A synergistic approach to protein crystallization: Combination of a fixed-arm carrier with surface entropy reduction Moon, Andrea F.; Mueller, Geoffrey A.; Zhong, Xuejun Protein Science https://doi.org/10.1002/pro.368	journal	January 2010
Crystal structure of the nuclear effector of Notch signaling, CSL, bound to DNA Kovall, Rhett A.; Hendrickson, Wayne A. The EMBO Journal, Vol. 23, Issue 17 https://doi.org/10.1038/sj.emboj.7600349	journal	August 2004
Structural Basis for Cooperativity in Recruitment of MAML Coactivators to Notch Transcription Complexes Nam, Yunsun; Sliz, Piotr; Song, Luyan Cell, Vol. 124, Issue 5 https://doi.org/10.1016/j.cell.2005.12.037	journal	March 2006
The contribution of Notch1 to nephron segmentation in the developing kidney is revealed in a sensitized Notch2 background and can be augmented by reducing Mint dosage Surendran, Kameswaran; Boyle, Scott; Barak, Hila Developmental Biology, Vol. 337, Issue 2 https://doi.org/10.1016/j.ydbio.2009.11.017	journal	January 2010
Structure and Function of the Su(H)-Hairless Repressor Complex, the Major Antagonist of Notch Signaling in Drosophila melanogaster Yuan, Zhenyu; Praxenthaler, Heiko; Tabaja, Nassif PLOS Biology, Vol. 14, Issue 7 https://doi.org/10.1371/journal.pbio.1002509	journal	July 2016
Chromatin Immunoprecipitation (ChIP) in Mouse T-cell Lines Giaimo, Benedetto Daniele; Ferrante, Francesca; Borggrefe, Tilman Journal of Visualized Experiments, Issue 124 https://doi.org/10.3791/55907	journal	June 2017
RITA, a novel modulator of Notch signalling, acts via nuclear export of RBP-J Wacker, Stephan Armin; Alvarado, Cristobal; von Wichert, Götz The EMBO Journal, Vol. 30, Issue 1 https://doi.org/10.1038/emboj.2010.289	journal	November 2010
Role of co-repressor genomic landscapes in shaping the Notch response Chan, Stephen K. K.; Cerda-Moya, Gustavo; Stojnic, Robert PLOS Genetics, Vol. 13, Issue 11 https://doi.org/10.1371/journal.pgen.1007096	journal	November 2017
Msx2-interacting nuclear target protein (Mint) deficiency reveals negative regulation of early thymocyte differentiation by Notch/RBP-J signaling Tsuji, Masayuki; Shinkura, Reiko; Kuroda, Kazuki Proceedings of the National Academy of Sciences, Vol. 104, Issue 5 https://doi.org/10.1073/pnas.0610520104	journal	January 2007
A conserved structural motif reveals the essential transcriptional repression function of Spen proteins and their role in developmental signaling Ariyoshi, Mariko; Schwabe, John W. R. Genes & Development, Vol. 17, Issue 15 https://doi.org/10.1101/gad.266203	journal	August 2003
Notch Signaling in Development, Tissue Homeostasis, and Disease Siebel, Chris; Lendahl, Urban Physiological Reviews, Vol. 97, Issue 4 https://doi.org/10.1152/physrev.00005.2017	journal	October 2017
PHENIX: a comprehensive Python-based system for macromolecular structure solution Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221 https://doi.org/10.1107/S0907444909052925	journal	January 2010
Regulation of Marginal Zone B Cell Development by MINT, a Suppressor of Notch/RBP-J Signaling Pathway Kuroda, Kazuki; Han, Hua; Tani, Shoichi Immunity, Vol. 18, Issue 2 https://doi.org/10.1016/S1074-7613(03)00029-3	journal	February 2003
Transcriptional Repression in the Notch Pathway: THERMODYNAMIC CHARACTERIZATION OF CSL-MINT (Msx2-INTERACTING NUCLEAR TARGET PROTEIN) COMPLEXES VanderWielen, Bradley D.; Yuan, Zhenyu; Friedmann, David R. Journal of Biological Chemistry, Vol. 286, Issue 17 https://doi.org/10.1074/jbc.M110.181156	journal	March 2011
SHARP is a novel component of the Notch/RBP-Jkappa signalling pathway Oswald, F. The EMBO Journal, Vol. 21, Issue 20 https://doi.org/10.1093/emboj/cdf549	journal	October 2002
The RRM Domain of MINT, a Novel Msx2 Binding Protein, Recognizes and Regulates the Rat Osteocalcin Promoter Newberry, Elizabeth P.; Latifi, Tammy; Towler, Dwight A. Biochemistry, Vol. 38, Issue 33 https://doi.org/10.1021/bi990967j	journal	July 1999
Sharp, an inducible cofactor that integrates nuclear receptor repression and activation Shi, Y. Genes & Development, Vol. 15, Issue 9 https://doi.org/10.1101/gad.871201	journal	May 2001
MolProbity: all-atom contacts and structure validation for proteins and nucleic acids Davis, I. W.; Leaver-Fay, A.; Chen, V. B. Nucleic Acids Research, Vol. 35, Issue Web Server https://doi.org/10.1093/nar/gkm216	journal	May 2007
Thermodynamic Analysis of the CSL·Notch Interaction Johnson, Scott E.; Ilagan, M. Xenia G.; Kopan, Raphael Journal of Biological Chemistry, Vol. 285, Issue 9 https://doi.org/10.1074/jbc.M109.019968	journal	February 2010
Notch signalling in context Bray, Sarah J. Nature Reviews Molecular Cell Biology, Vol. 17, Issue 11 https://doi.org/10.1038/nrm.2016.94	journal	August 2016
The Canonical Notch Signaling Pathway: Structural and Biochemical Insights into Shape, Sugar, and Force Kovall, Rhett A.; Gebelein, Brian; Sprinzak, David Developmental Cell, Vol. 41, Issue 3 https://doi.org/10.1016/j.devcel.2017.04.001	journal	May 2017
Quantitative Dissection of the Notch:CSL Interaction: Insights into the Notch-mediated Transcriptional Switch Lubman, Olga Y.; Ilagan, Ma. Xenia G.; Kopan, Raphael Journal of Molecular Biology, Vol. 365, Issue 3 https://doi.org/10.1016/j.jmb.2006.09.071	journal	January 2007
A phospho-dependent mechanism involving NCoR and KMT2D controls a permissive chromatin state at Notch target genes Oswald, Franz; Rodriguez, Patrick; Giaimo, Benedetto Daniele Nucleic Acids Research, Vol. 44, Issue 10 https://doi.org/10.1093/nar/gkw105	journal	February 2016
Dynamic binding of RBPJ is determined by Notch signaling status Castel, D.; Mourikis, P.; Bartels, S. J. J. Genes & Development, Vol. 27, Issue 9 https://doi.org/10.1101/gad.211912.112	journal	May 2013
The Notch signaling pathway: Transcriptional regulation at Notch target genes Borggrefe, T.; Oswald, F. Cellular and Molecular Life Sciences, Vol. 66, Issue 10 https://doi.org/10.1007/s00018-009-8668-7	journal	January 2009
From Fly Wings to Targeted Cancer Therapies: A Centennial for Notch Signaling Ntziachristos, Panagiotis; Lim, Jing Shan; Sage, Julien Cancer Cell, Vol. 25, Issue 3 https://doi.org/10.1016/j.ccr.2014.02.018	journal	March 2014
Hairless: the ignored antagonist of the Notch signalling pathway: Hairless: ignored antagonist of the Notch pathway Maier, Dieter Hereditas, Vol. 143, Issue 2006 https://doi.org/10.1111/j.2007.0018-0661.01971.x	journal	November 2006
Inference of Macromolecular Assemblies from Crystalline State Krissinel, Evgeny; Henrick, Kim Journal of Molecular Biology, Vol. 372, Issue 3 https://doi.org/10.1016/j.jmb.2007.05.022	journal	September 2007
Mutational and Energetic Studies of Notch1 Transcription Complexes Del Bianco, Cristina; Aster, Jon C.; Blacklow, Stephen C. Journal of Molecular Biology, Vol. 376, Issue 1 https://doi.org/10.1016/j.jmb.2007.11.061	journal	February 2008
RBPJ/CBF1 interacts with L3MBTL3/MBT1 to promote repression of Notch signaling via histone demethylaseKDM1A/LSD1 Xu, Tao; Park, Sung‐Soo; Giaimo, Benedetto Daniele The EMBO Journal, Vol. 36, Issue 21 https://doi.org/10.15252/embj.201796525	journal	October 2017
Crystal Structure of the CSL-Notch-Mastermind Ternary Complex Bound to DNA Wilson, Jeffrey J.; Kovall, Rhett A. Cell, Vol. 124, Issue 5 https://doi.org/10.1016/j.cell.2006.01.035	journal	March 2006
Conservation of the Notch antagonist Hairless in arthropods: functional analysis of the crustacean Daphnia pulex Hairless gene Zehender, Ariella; Bayer, Melanie; Bauer, Milena Development Genes and Evolution, Vol. 227, Issue 5 https://doi.org/10.1007/s00427-017-0593-4	journal	August 2017
Phaser crystallographic software McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D. Journal of Applied Crystallography, Vol. 40, Issue 4 https://doi.org/10.1107/S0021889807021206	journal	July 2007
Exploiting structure similarity in refinement: automated NCS and target-structure restraints in BUSTER Smart, Oliver S.; Womack, Thomas O.; Flensburg, Claus Acta Crystallographica Section D Biological Crystallography, Vol. 68, Issue 4 https://doi.org/10.1107/S0907444911056058	journal	March 2012
Characterization of CSL (CBF-1, Su(H), Lag-1) Mutants Reveals Differences in Signaling Mediated by Notch1 and Notch2 Yuan, Zhenyu; Friedmann, David R.; VanderWielen, Bradley D. Journal of Biological Chemistry, Vol. 287, Issue 42 https://doi.org/10.1074/jbc.M112.403287	journal	October 2012
The Notch Transcription Activation Complex Makes Its Move Barrick, Doug; Kopan, Raphael Cell, Vol. 124, Issue 5 https://doi.org/10.1016/j.cell.2006.02.028	journal	March 2006
Keeping Notch Target Genes off: A CSL Corepressor Caught in the Act Borggrefe, Tilman; Oswald, Franz Structure, Vol. 22, Issue 1 https://doi.org/10.1016/j.str.2013.12.007	journal	January 2014
Structure-function analysis of RBP-J-interacting and tubulin-associated (RITA) reveals regions critical for repression of Notch target genes Tabaja, Nassif; Yuan, Zhenyu; Oswald, Franz Journal of Biological Chemistry, Vol. 292, Issue 25 https://doi.org/10.1074/jbc.m117.791707	journal	June 2017
RAM-induced Allostery Facilitates Assembly of a Notch Pathway Active Transcription Complex Friedmann, David R.; Wilson, Jeffrey J.; Kovall, Rhett A. Journal of Biological Chemistry, Vol. 283, Issue 21 https://doi.org/10.1074/jbc.m709501200	journal	April 2008
LIM Protein KyoT2 Negatively Regulates Transcription by Association with the RBP-J DNA-Binding Protein Taniguchi, Yoshihito; Furukawa, Takahisa; Tun, Tin Molecular and Cellular Biology, Vol. 18, Issue 1 https://doi.org/10.1128/mcb.18.1.644	journal	January 1998
Setting the Stage for Notch: The Drosophila Su(H)-Hairless Repressor Complex Borggrefe, Tilman; Oswald, Franz PLOS Biology, Vol. 14, Issue 7 https://doi.org/10.1371/journal.pbio.1002524	journal	July 2016
MolProbity: all-atom contacts and structure validation for proteins and nucleic acids B., Chen, Vincent; C., Richardson, David; S., Richardson, Jane The University of North Carolina at Chapel Hill University Libraries https://doi.org/10.17615/m2an-p252	text	January 2007
Role of co-repressor genomic landscapes in shaping the Notch response. Chan, Stephen KK; Cerda-Moya, Gustavo; Stojnic, Robert Public Library of Science (PLoS) https://doi.org/10.17863/cam.20237	journalarticle	January 2017
PHENIX: a comprehensive Python-based system for macromolecular structure solution. Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.45787	text	January 2010

Figures / Tables (4)

Similar Records

Crystal Structure of the CSL-Notch-Mastermind Ternary Complex Bound to DNA

Journal Article · Sun Jan 01 00:00:00 EST 2006 · Cell · OSTI ID:1569868

Wilson, J; Kovall, R

Corepressors of agonist-bound nuclear receptors

Journal Article · Sat Sep 15 00:00:00 EDT 2007 · Toxicology and Applied Pharmacology · OSTI ID:1569868

Gurevich, Igor; Flores, Anthony M

Structure and Function of the Su(H)-Hairless Repressor Complex, the Major Antagonist of Notch Signaling in Drosophila melanogaster

Journal Article · Tue Jul 12 00:00:00 EDT 2016 · PLoS Biology (Online) · OSTI ID:1569868

Yuan, Zhenyu; Praxenthaler, Heiko; Tabaja, Nassif; +5 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
notch signaling
CSL
RBPJ
SHARP
MINT
SPEN
transcriptional regulation
signal transduction
x-ray crystallography
isothermal titration calorimetry

Title: Structural and Functional Studies of the RBPJ-SHARP Complex Reveal a Conserved Corepressor Binding Site

Citation Formats

References (47)

Figures / Tables (4)

Similar Records

Related Subjects