TY - JOUR
T1 - Conformational locking upon cooperative assembly of notch transcription complexes
AU - Choi, Sung Hee
AU - Wales, Thomas E.
AU - Nam, Yunsun
AU - O'Donovan, Daniel J.
AU - Sliz, Piotr
AU - Engen, John R.
AU - Blacklow, Stephen C.
N1 - Funding Information:
We thank Jon Aster, Kelly Arnett, and Wendy Gordon for helpful discussions and critical reading of the manuscript. We also thank two anonymous reviewers for their thoughtful comments and suggestions for revision. This work was supported by NIH grants R01 CA092433, P01 CA119070, R01 GM086507, NSF grant RCN 0639193, a research collaboration with the Waters Corporation, and a Leukemia and Lymphoma Society Special Center of Research. This is contribution 988 from the Barnett Institute.
PY - 2012/2/8
Y1 - 2012/2/8
N2 - The Notch intracellular domain (NICD) forms a transcriptional activation complex with the DNA-binding factor CSL and a transcriptional co-activator of the Mastermind family (MAML). The "RAM" region of NICD recruits Notch to CSL, facilitating the binding of MAML at the interface between the ankyrin (ANK) repeat domain of NICD and CSL. Here, we report the X-ray structure of a human MAML1/RAM/ANK/CSL/DNA complex, and probe changes in component dynamics upon stepwise assembly of a MAML1/NICD/CSL complex using HX-MS. Association of CSL with NICD exerts remarkably little effect on the exchange kinetics of the ANK domain, whereas MAML1 binding greatly retards the exchange kinetics of ANK repeats 2-3. These exchange patterns identify critical features contributing to the cooperative assembly of Notch transcription complexes (NTCs), highlight the importance of MAML recruitment in rigidifying the ANK domain and stabilizing its interface with CSL, and rationalize the requirement for MAML1 in driving cooperative dimerization of NTCs on paired-site DNA.
AB - The Notch intracellular domain (NICD) forms a transcriptional activation complex with the DNA-binding factor CSL and a transcriptional co-activator of the Mastermind family (MAML). The "RAM" region of NICD recruits Notch to CSL, facilitating the binding of MAML at the interface between the ankyrin (ANK) repeat domain of NICD and CSL. Here, we report the X-ray structure of a human MAML1/RAM/ANK/CSL/DNA complex, and probe changes in component dynamics upon stepwise assembly of a MAML1/NICD/CSL complex using HX-MS. Association of CSL with NICD exerts remarkably little effect on the exchange kinetics of the ANK domain, whereas MAML1 binding greatly retards the exchange kinetics of ANK repeats 2-3. These exchange patterns identify critical features contributing to the cooperative assembly of Notch transcription complexes (NTCs), highlight the importance of MAML recruitment in rigidifying the ANK domain and stabilizing its interface with CSL, and rationalize the requirement for MAML1 in driving cooperative dimerization of NTCs on paired-site DNA.
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U2 - 10.1016/j.str.2011.12.011
DO - 10.1016/j.str.2011.12.011
M3 - Article
C2 - 22325781
AN - SCOPUS:84863012561
SN - 0969-2126
VL - 20
SP - 340
EP - 349
JO - Structure
JF - Structure
IS - 2
ER -