Cadherin cell adhesion molecules link the cytoskeletons of adjacent cells together in solid tissues. In adherens junctions, the catenins link cadherins to the actin-based cytoskeleton, and in desmosomes an analogous protein assembly links cadherins to intermediate filaments. Assembly of adherens junction is a critical step in the development of cell and tissue morphology, and loss of adherens junction-based cell adhesion is a hallmark of metastasizing cells. Linkage to the intermediate filament system is essential for the mechanical strength and integrity of tissues such as the skin and the heart, and defects in desmosome assembly are responsible for a number of severe skin blistering diseases. The adherens junction protein beta-catenin also serves as a transcriptional coactivator in the Wnt signaling pathway that controls cell fate determination during embryogenesis. Mutations of Wnt signaling components are responsible for many cancers. This proposal aims to develop a biophysical understanding of cell junction architecture and assembly, and also the interactions of beta-catenin in Wnt signaling, using x-ray crystallography to determine structures and physical biochemical methods to determine binding stoichiometries, affinities, and kinetics. 1. The following structures will be determined: a) phosphorylated E-cadherin/beta-catenin complex; b) a ternary complex of E-cadherin, beta-catenin, and alpha-catenin; c) the actin-binding domain of a-catenin; and d) full-length alpha-catenin. The affinity and kinetics of the beta-catenin/phosphorylated E-cadherin interaction will be measured. 2. The structures of domains from desmoplakin, which links the cadherin/plakoglobin complex to intermediate filaments, will be determined to understand how this protein interacts with intermediate filaments and with the cadherin/plakoglobin complex. Complexes of cadherins with plakoglobin will also be studied. 3. The binding properties of different sets of beta-catenin binding repeats of the tumor suppressor APC, both unmodified and phosphorylated, will be determined, and complexes with beta-catenin and Axin will be crystallized. The structure of a ternary complex of beta-catenin, the transcription factor LEF-1, and a cognate DNA sequence will be determined.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM056169-05
Application #
6382801
Study Section
Physiological Chemistry Study Section (PC)
Program Officer
Flicker, Paula F
Project Start
1997-08-01
Project End
2005-07-31
Budget Start
2001-08-01
Budget End
2002-07-31
Support Year
5
Fiscal Year
2001
Total Cost
$309,440
Indirect Cost
Name
Stanford University
Department
Biology
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Kang, Hyunook; Weiss, Thomas M; Bang, Injin et al. (2016) Structure of the Intermediate Filament-Binding Region of Desmoplakin. PLoS One 11:e0147641
Dickinson, Daniel J; Nelson, W James; Weis, William I (2015) Studying epithelial morphogenesis in Dictyostelium. Methods Mol Biol 1189:267-81
Choi, Hee-Jung; Loveless, Timothy; Lynch, Allison M et al. (2015) A conserved phosphorylation switch controls the interaction between cadherin and ?-catenin in vitro and in vivo. Dev Cell 33:82-93
Choi, Hee-Jung; Weis, William I (2015) Purification, crystallization and preliminary X-ray diffraction studies of the ?-catenin homolog HMP-2 from Caenorhabditis elegans. Acta Crystallogr F Struct Biol Commun 71:272-6
Stamos, Jennifer L; Chu, Matthew Ling-Hon; Enos, Michael D et al. (2014) Structural basis of GSK-3 inhibition by N-terminal phosphorylation and by the Wnt receptor LRP6. Elife 3:e01998
Buckley, Craig D; Tan, Jiongyi; Anderson, Karen L et al. (2014) Cell adhesion. The minimal cadherin-catenin complex binds to actin filaments under force. Science 346:1254211
Pokutta, Sabine; Choi, Hee-Jung; Ahlsen, Goran et al. (2014) Structural and thermodynamic characterization of cadherin·?-catenin·?-catenin complex formation. J Biol Chem 289:13589-601
Chodaparambil, Jayanth V; Pate, Kira T; Hepler, Margretta R D et al. (2014) Molecular functions of the TLE tetramerization domain in Wnt target gene repression. EMBO J 33:719-31
Weis, William I; Nelson, W James; Dickinson, Daniel J (2013) Evolution and cell physiology. 3. Using Dictyostelium discoideum to investigate mechanisms of epithelial polarity. Am J Physiol Cell Physiol 305:C1091-5
Nelson, W James; Dickinson, Daniel J; Weis, William I (2013) Roles of cadherins and catenins in cell-cell adhesion and epithelial cell polarity. Prog Mol Biol Transl Sci 116:3-23

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