The goal of this research is to determine how the assembly dynamics and architecture of the actin cytoskeleton are controlled by formins and the mechanisms regulating their activities in vivo. We are studying this question in budding yeast, where the formins Bni1 and Bnr1 assemble actin 'cables'that play an essential role in polarized cell growth. Our lab recently discovered four novel modes of formin regulation mediated by the polarity factors Bud6, Bud14, Smy1, and Hof1. Remarkably, each of these proteins binds to the formin-homology FH2 domain of Bnr1, but has distinct effects on Bnr1 activity in vitro and distinct Bnr1-dependent mutant actin cable phenotypes in vivo. Further, we have identified novel in vivo ligands of Bud6 and Bud14 that function with them in regulating Bnr1-mediated actin cable assembly. The proposed research will define the cellular functions and mechanisms of these proteins, and how their combined effects coordinate the proper assembly of actin cables with a characteristic length, architecture, and dynamics that is tailored to their function. This work will provide a deeper understanding of the molecular activities and interactions that underlie cell polarity and morphogenesis. The project uses a multi-disciplinary approach, combining genetics, live-cell imaging, biochemistry, and novel multi-wavelength single molecule TIRF in vitro microscopy.
The Aims are to: (1) Elucidate the specific roles and mechanisms of Bud6 in regulating Bnr1- mediated actin cable assembly;(2) Test the hypothesis that Bud14 and Smy1 provide distinct modes of formin temporal regulation required for maintaining actin cable length, dynamics, and architecture;and (3) Determine how the functions of multiple FH2-binding regulators are coordinated in vitro and in vivo.

Public Health Relevance

The proper function of human tissues and organs depends on cells within those tissues maintaining their sense of direction (or polarity), dividing properly, ad maintaining their characteristic shapes and functions. Our work addresses a family of proteins (formins) that play vital roles in controlling cell shape, polarity, and division. By determining hw formins are regulated on a molecular and cellular level, we anticipate that this research will provide new insights into the underlying mechanisms of tumorigenesis, neurodegeneration, and developmental disorders such as limb deformities and hearing loss, which result from misregulated cell shape and division.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM083137-07S1
Application #
8790310
Study Section
Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Gindhart, Joseph G
Project Start
2008-03-01
Project End
2016-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
7
Fiscal Year
2014
Total Cost
$38,607
Indirect Cost
$9,136
Name
Brandeis University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
Garabedian, Mikael V; Stanishneva-Konovalova, Tatiana; Lou, Chenyu et al. (2018) Integrated control of formin-mediated actin assembly by a stationary inhibitor and a mobile activator. J Cell Biol 217:3512-3530
Henty-Ridilla, Jessica L; Juanes, M Angeles; Goode, Bruce L (2017) Profilin Directly Promotes Microtubule Growth through Residues Mutated in Amyotrophic Lateral Sclerosis. Curr Biol 27:3535-3543.e4
Sokolova, Olga S; Chemeris, Angelina; Guo, Siyang et al. (2017) Structural Basis of Arp2/3 Complex Inhibition by GMF, Coronin, and Arpin. J Mol Biol 429:237-248
Shekhar, Shashank (2017) Microfluidics-Assisted TIRF Imaging to Study Single Actin Filament Dynamics. Curr Protoc Cell Biol 77:12.13.1-12.13.24
Henty-Ridilla, Jessica L; Rankova, Aneliya; Eskin, Julian A et al. (2016) Accelerated actin filament polymerization from microtubule plus ends. Science 352:1004-9
Alioto, Salvatore L; Garabedian, Mikael V; Bellavance, Danielle R et al. (2016) Tropomyosin and Profilin Cooperate to Promote Formin-Mediated Actin Nucleation and Drive Yeast Actin Cable Assembly. Curr Biol 26:3230-3237
Mohapatra, Lishibanya; Goode, Bruce L; Jelenkovic, Predrag et al. (2016) Design Principles of Length Control of Cytoskeletal Structures. Annu Rev Biophys 45:85-116
Eskin, Julian A; Rankova, Aneliya; Johnston, Adam B et al. (2016) Common formin-regulating sequences in Smy1 and Bud14 are required for the control of actin cable assembly in vivo. Mol Biol Cell 27:828-37
Bombardier, Jeffrey P; Eskin, Julian A; Jaiswal, Richa et al. (2015) Single-molecule visualization of a formin-capping protein 'decision complex' at the actin filament barbed end. Nat Commun 6:8707
Mohapatra, Lishibanya; Goode, Bruce L; Kondev, Jane (2015) Antenna Mechanism of Length Control of Actin Cables. PLoS Comput Biol 11:e1004160

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