The goal of this research is to determine how the dynamics of the actin and microtubule cytoskeletons are coordinated by a group of three interacting mammalian proteins: APC, Dia1, and EB1. This work will define the functions and mechanisms of these proteins, and provide a deeper understanding of the activities and interactions that underlie such processes as cell migration and morphogenesis. This project uses bulk biochemical experiments combined with a novel multi-wavelength single molecule biophysics method tailored to elucidate the mechanisms of complex, multi-component regulatory systems in vitro. In addition, the mechanisms deduced from the experiments in vitro will be tested in vivo to verify that they are important for relevant biological functions of these proteins in living cells (e.g., directed cell migration).
The Specific Aims are: (1) Test two key hypotheses about the mechanism by which Dia1 and APC synergize in promoting actin assembly, involving formation of proposed physical complexes among components;(2) Test the additional hypothesis that Dia1 and APC synergize to stimulate actin assembly by a "rocket launcher" mechanism;and (3) Define the mechanisms by which EB1 alone and EB1 plus microtubules regulate and/or organize APC/Dia-induced actin assembly.

Public Health Relevance

This grant is an investigation of the organization and regulation of actin and microtubule cytoskeletons by the combined effects of three proteins: APC, Dia1, and EB1. APC (Adenomatous polyposis coli) is a tumor suppressor. Mutation of the human Apc gene is considered to be an early step in the progression of over 80% of colorectal cancers, inherited and sporadic. In addition, fusion of the EB1 and MLL genes can lead to acute lymphoblastic leukemia. We anticipate that this research will provide new insights into the underlying mechanisms of tumorigenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM098143-02
Application #
8454423
Study Section
Special Emphasis Panel (ZRG1-BCMB-B (02))
Program Officer
Gindhart, Joseph G
Project Start
2012-05-01
Project End
2016-02-29
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
2
Fiscal Year
2013
Total Cost
$311,374
Indirect Cost
$118,374
Name
Brandeis University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
Daou, Pascale; Hasan, Salma; Breitsprecher, Dennis et al. (2014) Essential and nonredundant roles for Diaphanous formins in cortical microtubule capture and directed cell migration. Mol Biol Cell 25:658-68
Smith, Benjamin A; Gelles, Jeff; Goode, Bruce L (2014) Single-molecule studies of actin assembly and disassembly factors. Methods Enzymol 540:95-117
Jaiswal, Richa; Breitsprecher, Dennis; Collins, Agnieszka et al. (2013) The formin Daam1 and fascin directly collaborate to promote filopodia formation. Curr Biol 23:1373-9
Jaiswal, Richa; Stepanik, Vince; Rankova, Aneliya et al. (2013) Drosophila homologues of adenomatous polyposis coli (APC) and the formin diaphanous collaborate by a conserved mechanism to stimulate actin filament assembly. J Biol Chem 288:13897-905