The goal of the work described in these studies is to elucidate how critical extracellular and intracellular domains control signaling by notch receptors. The extracellular domains of all notches contain three iterated LIN12 repeats, which maintain notch receptors in their resting state prior to ligand binding. Ligand-induced proteolysis releases the intracellular portion of notch (ICN) and allow it to translocate to the nucleus, where its RAM and ankyrin-repeat domains interact with two additional factors, RBP-JK and mastermind (mami 1) to form a transcriptionally active ternary complex.
The specific aims of this research plan, which will provide important new insights into the structure and function of the LIN12 and ankyrin repeat domains of notch, are to: ? ? 1. Determine how LIN12 modules prevent ligand-independent activation of notch receptors. (a) Determine high-resolution solution structures of key modules and module pairs from a Notch LIN12 domain, analyzing the structural interactions of these modules with each other and with neighboring domains. (b) Identify contacts between the LIN12 domain and the NTM extracellular stub that maintain the integrity of the Notch heterodimer, and (c) Test the functional importance of specific intermodular and LIN12-NTM contacts. ? ? 2. Determine the essential domains and residues required for assembly and function of thenotch/RBP-Jkappa/maml1 signaling complex. (a) Determine the minimal protein domains required for reconstitution of an ICN-mam11-RBP-Jkappa--DNA complex. (b) Identify contacts that contribute to energetic stabilization of ICN complexes and determine how RBP and RAMANK cooperate to recruit mamli to form a ternary complex on DNA. (c) Test the function of specific contacts responsible for stabilizing the ternary DNA-binding complex.
|Aster, Jon C; Pear, Warren S; Blacklow, Stephen C (2017) The Varied Roles of Notch in Cancer. Annu Rev Pathol 12:245-275|
|Seegar, Tom C M; Killingsworth, Lauren B; Saha, Nayanendu et al. (2017) Structural Basis for Regulated Proteolysis by the ?-Secretase ADAM10. Cell 171:1638-1648.e7|
|Durzynska, Izabela; Xu, Xiang; Adelmant, Guillaume et al. (2017) STK40 Is a Pseudokinase that Binds the E3 Ubiquitin Ligase COP1. Structure 25:287-294|
|Severson, Eric; Arnett, Kelly L; Wang, Hongfang et al. (2017) Genome-wide identification and characterization of Notch transcription complex-binding sequence-paired sites in leukemia cells. Sci Signal 10:|
|Ryan, Russell J H; Petrovic, Jelena; Rausch, Dylan M et al. (2017) A B Cell Regulome Links Notch to Downstream Oncogenic Pathways in Small B Cell Lymphomas. Cell Rep 21:784-797|
|McMillan, Brian J; Tibbe, Christine; Drabek, Andrew A et al. (2017) Structural Basis for Regulation of ESCRT-III Complexes by Lgd. Cell Rep 19:1750-1757|
|McMillan, Brian J; Zimmerman, Brandon; Egan, Emily D et al. (2017) Structure of human POFUT1, its requirement in ligand-independent oncogenic Notch signaling, and functional effects of Dowling-Degos mutations. Glycobiology 27:777-786|
|Blacklow, Stephen C (2017) Signal Transduction: Notch catches a Jagged edge. Nat Chem Biol 13:570-571|
|Zimmerman, Brandon; Kelly, Brendan; McMillan, Brian J et al. (2016) Crystal Structure of a Full-Length Human Tetraspanin Reveals a Cholesterol-Binding Pocket. Cell 167:1041-1051.e11|
|McMillan, Brian J; Tibbe, Christine; Jeon, Hyesung et al. (2016) Electrostatic Interactions between Elongated Monomers Drive Filamentation of Drosophila Shrub, a Metazoan ESCRT-III Protein. Cell Rep 16:1211-1217|
Showing the most recent 10 out of 50 publications