The continued goal of this Initiative is to determine the high resolution structures of two major migration organelles, adhesions and protrusions. Our approach is to combine structural methods that range from correlative light and high resolution electron microscopy to X-ray and NMR. As part of this effort, we are developing a new imaging concept that allows us to capture by light microscopy the spatial-temporal aspects of migration in living cells and to visualize the same structures using high resolution electron microscopy - 3D correlative light and electron microscopy (""""""""top down approaches""""""""). High resolution structures of individual proteins and their detailed interactions with binding partners obtained by NMR, X-ray or tomography (""""""""bottom up approaches"""""""") can then be computationally docked into the three-dimensional maps of the complexes. In the first phase of the Consortium, this paradigm was applied to the structure of the actin network in protrusions, providing 3D structures of Arp2/3 mediated actin branches and the structural basis for Arp2/3 activation. We also determined the ultrastructure of the lamellipodia of living cells using our new 3D correlative EM and speckle microscopy and determined the vinculin binding site on a-actinin in the presence of integrin. A unified framework for segmentation and analysis of 3-D data was developed to enable the identification of assemblies in 3-D structures.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54GM064346-10
Application #
8121487
Study Section
Special Emphasis Panel (ZGM1)
Project Start
Project End
2014-07-31
Budget Start
2010-08-01
Budget End
2013-07-31
Support Year
10
Fiscal Year
2010
Total Cost
$167,977
Indirect Cost
Name
University of Virginia
Department
Type
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Hu, Guiqing; Taylor, Dianne W; Liu, Jun et al. (2018) Identification of interfaces involved in weak interactions with application to F-actin-aldolase rafts. J Struct Biol 201:199-209
Kubow, Kristopher E; Shuklis, Victoria D; Sales, Dominic J et al. (2017) Contact guidance persists under myosin inhibition due to the local alignment of adhesions and individual protrusions. Sci Rep 7:14380
Gallegos, Lisa Leon; Ng, Mei Rosa; Sowa, Mathew E et al. (2016) A protein interaction map for cell-cell adhesion regulators identifies DUSP23 as a novel phosphatase for ?-catenin. Sci Rep 6:27114
Al-Dimassi, Saleh; Salloum, Gilbert; Saykali, Bechara et al. (2016) Targeting the MAP kinase pathway in astrocytoma cells using a recombinant anthrax lethal toxin as a way to inhibit cell motility and invasion. Int J Oncol 48:1913-20
Juanes-Garcia, Alba; Chapman, Jessica R; Aguilar-Cuenca, Rocio et al. (2015) A regulatory motif in nonmuscle myosin II-B regulates its role in migratory front-back polarity. J Cell Biol 209:23-32
Gao, Runchi; Zhao, Siwei; Jiang, Xupin et al. (2015) A large-scale screen reveals genes that mediate electrotaxis in Dictyostelium discoideum. Sci Signal 8:ra50
Dai, Aguang; Ye, Feng; Taylor, Dianne W et al. (2015) The Structure of a Full-length Membrane-embedded Integrin Bound to a Physiological Ligand. J Biol Chem 290:27168-75
Hanna, Samer; Khalil, Bassem; Nasrallah, Anita et al. (2014) StarD13 is a tumor suppressor in breast cancer that regulates cell motility and invasion. Int J Oncol 44:1499-511
Itano, Michelle S; Graus, Matthew S; Pehlke, Carolyn et al. (2014) Super-resolution imaging of C-type lectin spatial rearrangement within the dendritic cell plasma membrane at fungal microbe contact sites. Front Phys 2:
Wong, Ming-Ching; Kennedy, William P; Schwarzbauer, Jean E (2014) Transcriptionally regulated cell adhesion network dictates distal tip cell directionality. Dev Dyn 243:999-1010

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