The long-term goal of this program project is to understand the molecular mechanisms that allow Arp2/3 complex to control actin filament assembly during endocytosis and cellular motility. The mechanism of action of Arp2/3 complex is so complicated that a team effort is required. Our group of four geographically dispersed laboratories combines x-ray crystallography, electron microscopy, image processing and molecular biology to address technically challenging questions. Structural insights from his ongoing collaboration will provide important clues about reaction mechanisms and also help to design experiments to measure kinetic and thermodynamic constants and to study actin assembly in live cells. Project 1 (Pollard) proposes a combination of biochemical and x-ray crystallographic experiments to obtain detailed structural information regarding conformational changes and macromolecular interactions of Arp2/3 complex. Project 2 (Li) proposes a combination of biophysical studies on interaction's of nucleation promoting factors with purified Arp2/3 complex and microscopic analysis of mouse cells lacking subunits of Arp2/3 complex. Project 3 (Hanein) proposes electron microscopy of purified Arp2/3 complex, actin filament branches and of actin networks in mouse cells with and without functional Arp2/3 complex. Project 4 (Volkmann) proposes computational methods to extract the maximal amount of information from spectroscopy experiments in projects 1 and 2 and the electron micrographs from project 3.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM066311-07
Application #
7821395
Study Section
Special Emphasis Panel (ZRG1-CB-D (40))
Program Officer
Gindhart, Joseph G
Project Start
2003-06-10
Project End
2014-04-30
Budget Start
2010-05-01
Budget End
2011-04-30
Support Year
7
Fiscal Year
2010
Total Cost
$1,450,243
Indirect Cost
Name
Yale University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Anderson, Karen L; Page, Christopher; Swift, Mark F et al. (2017) Nano-scale actin-network characterization of fibroblast cells lacking functional Arp2/3 complex. J Struct Biol 197:312-321
Wang, Pei-Shan; Chou, Fu-Sheng; Ramachandran, Sreekumar et al. (2016) Crucial roles of the Arp2/3 complex during mammalian corticogenesis. Development 143:2741-52
Tee, Yee Han; Shemesh, Tom; Thiagarajan, Visalatchi et al. (2015) Cellular chirality arising from the self-organization of the actin cytoskeleton. Nat Cell Biol 17:445-57
Jurgenson, Christopher T; Pollard, Thomas D (2015) Crystals of the Arp2/3 complex in two new space groups with structural information about actin-related protein 2 and potential WASP binding sites. Acta Crystallogr F Struct Biol Commun 71:1161-8
Li, Yongchao; Wang, Pei-Shan; Lucas, George et al. (2015) ARP2/3 complex is required for directional migration of neural stem cell-derived oligodendrocyte precursors in electric fields. Stem Cell Res Ther 6:41
Suraneni, Praveen; Fogelson, Ben; Rubinstein, Boris et al. (2015) A mechanism of leading-edge protrusion in the absence of Arp2/3 complex. Mol Biol Cell 26:901-12
Page, Christopher; Hanein, Dorit; Volkmann, Niels (2015) Accurate membrane tracing in three-dimensional reconstructions from electron cryotomography data. Ultramicroscopy 155:20-6
Volkmann, Niels (2014) The joys and perils of flexible fitting. Adv Exp Med Biol 805:137-55
Volkmann, Niels; Page, Christopher; Li, Rong et al. (2014) Three-dimensional reconstructions of actin filaments capped by Arp2/3 complex. Eur J Cell Biol 93:179-83
Yi, Kexi; Rubinstein, Boris; Li, Rong (2013) Symmetry breaking and polarity establishment during mouse oocyte maturation. Philos Trans R Soc Lond B Biol Sci 368:20130002

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