Our lab is focused on the question of how cells dynamically assemble and reorganize their actin cytoskeletons to govern cell shape, cell movement, and cell division. Every cell type has a unique architecture tailored to its physiological functions, which is defined in large part by its cytoskeleton. The cytoskeleton is a vast system of interconnected polymeric tubes and fibers that creates a dynamic scaffold used to produce polarity and compartmentalization, and to generate force. The primary aim of our research is to understand how cells bring about rapid and precise rearrangements of their actin polymer networks, transforming cell shape and function.
Actin filament networks are complex, self-assembling biological machines built from hundreds of distinct components and moving parts. We have been dissecting the inner workings of these force-producing machines, analyzing the cellular functions and biochemical mechanisms of actin regulatory proteins in order to understand how this machinery collectively orchestrates the assembly, reorganization, and disassembly of entire actin networks. Our goal is to obtain a highly mechanistic and quantitative view of these events.
We use two different in vivo systems: (a) the budding yeast S. cerevisiae, which offers advanced genetic analyses; and (b) mammalian fibroblasts and primary cardiomyocytes, which offer superior cytology and enable us to study actin regulation underlying cell motility and muscle contraction. Two fundamental questions are being addressed in the lab:
- How is the rapid assembly and disassembly of actin networks governed? To answer this question, we are studying the roles of crucial actin regulatory proteins that alter polymer dynamics, e.g. formins, Arp2/3 complex, and ADF/cofilin (figure 1). We have also identified novel binding partners of these regulators that have surprising new activities. This work includes the characterization of: (a) yeast and mammalian formins, (b) formin-binding proteins (e.g. APC, Bud6, Bud14 and Smy1), (c) WASp-Arp2/3 complex binding partners (e.g. Abp1, Syp1, coronin, and GMF), and (d) the actin disassembly and turnover machinery (e.g. Aip1, coronin, twinfilin, and Srv2/CAP).
- How is the regulation of actin dynamics coordinated with regulation of other cytoskeletal polymer systems (microtubules, septins, and intermediate filaments)? Our recent work in this area addresses functional cross-talk between the actin and microtubule cytoskeletons regulated by a triad of mammalian proteins that physically associate: (a) the tumor suppressor protein Adenomatous polyposis coli (APC), (b) the formin Dia1, and (c) the microtubule end-binding protein EB
To answer the questions above, we are taking a multidisciplinary approach combining genetics, biochemistry, structural biology, and live cell imaging. In vitro analyses include quantitative fluorescence-based kinetic assays, time lapse TIRF microscopy on individual actin filaments, single molecule analysis on actin regulatory proteins, and reconstituted actin-based bead motility assays in cell extracts and purified systems. In vivo analyses include forward and reverse genetic screens, RNAi silencing, live-cell imaging to track cytoskeletal regulatory protein dynamics and polymer dynamics, and electron microscopy (figure 2).
Recent Publications (2010-present)
Ydenberg CA, Padrick SB, Sweeney MO, Gandhi M, Sokolova O, Goode BL. GMF severs actin-Arp2/3 complex branch junctions by a cofilin-like mechanism. Curr. Biol. 2013;23(12):1037-45. [abstract] [full text in PubMed Central].
Smith BA, Padrick SB, Doolittle LK, Daugherty-Clarke K, Correa IR, Jr., Xu MQ, Goode BL, Rosen MK, Gelles J. Three-color single molecule imaging shows WASP detachment from Arp2/3 complex triggers actin filament branch formation. eLife. 2013;2(0):e01008. [abstract].
Smith BA, Daugherty-Clarke K, Goode BL, Gelles J. Pathway of actin filament branch formation by Arp2/3 complex revealed by single-molecule imaging. Proc Natl Acad Sci U S A. 2013;110(4):1285-90. [abstract] [full text in PubMed Central].
Jaiswal R, Stepanik V, Rankova A, Molinar O, Goode BL, McCartney BM. Drosophila Homologues of Adenomatous Polyposis Coli (APC) and the Formin Diaphanous Collaborate by a Conserved Mechanism to Stimulate Actin Filament Assembly. J Biol Chem. 2013;288(19):13897-905. [abstract] [full text in PubMed Central].
Jaiswal R, Breitsprecher D, Collins A, Correa IR, Jr., Xu MQ, Goode BL. The formin daam1 and fascin directly collaborate to promote filopodia formation. Curr Biol. 2013;23(14):1373-9. [abstract] [full text in PubMed Central].
Graziano BR, Jonasson EM, Pullen JG, Gould CJ, Goode BL. Ligand-induced activation of a formin-NPF pair leads to collaborative actin nucleation. J Cell Biol. 2013;201(4):595-611. [abstract] [full text in PubMed Central].
Chaudhry F, Breitsprecher D, Little K, Sharov G, Sokolova O, Goode BL. Srv2/cyclase-associated protein forms hexameric shurikens that directly catalyze actin filament severing by cofilin. Mol Biol Cell. 2013;24(1):31-41. [abstract] [full text in PubMed Central].
Breitsprecher D, Goode BL. Formins at a glance. J Cell Sci. 2013;126(Pt 1):1-7. [abstract] [full text in PubMed Central].
Tu D, Graziano BR, Park E, Zheng W, Li Y, Goode BL, Eck MJ. Structure of the formin-interaction domain of the actin nucleation-promoting factor Bud6. Proc Natl Acad Sci U S A. 2012;109(50):E3424-33. [abstract] [full text in PubMed Central].
Maiti S, Michelot A, Gould C, Blanchoin L, Sokolova O, Goode BL. Structure and activity of full-length formin mDia1. Cytoskeleton (Hoboken). 2012;69(6):393-405. [abstract] [full text in PubMed Central].
Breitsprecher D, Jaiswal R, Bombardier JP, Gould CJ, Gelles J, Goode BL. Rocket launcher mechanism of collaborative actin assembly defined by single-molecule imaging. Science. 2012;336(6085):1164-8. [abstract].
Ydenberg CA, Smith BA, Breitsprecher D, Gelles J, Goode BL. Cease-fire at the leading edge: new perspectives on actin filament branching, debranching, and cross-linking. Cytoskeleton (Hoboken). 2011;68(11):596-602. [abstract] [full text in PubMed Central].
Tu D, Li Y, Song HK, Toms AV, Gould CJ, Ficarro SB, Marto JA, Goode BL, Eck MJ. Crystal Structure of a Coiled-Coil Domain from Human ROCK I. PLoS One. 2011;6(3):e18080. [abstract] [full text in PubMed Central].
Huang W, Ghisletti S, Saijo K, Gandhi M, Aouadi M, Tesz GJ, Zhang DX, Yao J, Czech MP, Goode BL, Rosenfeld MG, Glass CK. Coronin 2A mediates actin-dependent de-repression of inflammatory response genes. Nature. 2011;470(7334):414-8. [abstract].
Graziano BR, Dupage AG, Michelot A, Breitsprecher D, Moseley JB, Sagot I, Blanchoin L, Goode BL. Mechanism and cellular function of Bud6 as an actin nucleation-promoting factor. Mol Biol Cell. 2011;22(21):4016-28. [abstract] [full text in PubMed Central].
Gould CJ, Maiti S, Michelot A, Graziano BR, Blanchoin L, Goode BL. The formin DAD domain plays dual roles in autoinhibition and actin nucleation. Curr Biol. 2011;21(5):384-90. [abstract] [full text in PubMed Central].
Chesarone-Cataldo M, Guerin C, Yu JH, Wedlich-Soldner R, Blanchoin L, Goode BL. The myosin passenger protein Smy1 controls actin cable structure and dynamics by acting as a formin damper. Dev Cell. 2011;21(2):217-30. [abstract] [full text in PubMed Central].
Breitsprecher D, Koestler SA, Chizhov I, Nemethova M, Mueller J, Goode BL, Small JV, Rottner K, Faix J. Cofilin cooperates with fascin to disassemble filopodial actin filaments. J Cell Sci. 2011;124(Pt 19):3305-18. [abstract].
Gandhi, M., M. Jangi, and B.L. Goode (2010). Identification of conserved surfaces of coronin required for actin binding and in vivo functions. J. Biol. Chem. 285:34899-908.
Okada, K., F. Bartoloni, A. Deaconescu, J.B. Moseley, Z. Dogic, N. Grigorieff, G.G. Gundersen, and B.L. Goode (2010). Adenomatous polyposis coli protein nucleates actin assembly and synergizes with the formin mDia1. J. Cell Biol. 189(7):1087-96. [abstract]
Gandhi, M., B. Smith, M. Bovellan, V. Paavilainen, K. Daugherty-Clarke, J. Gelles, P. Lappalainen, and B.L. Goode (2010). GMF is a cofilin homologue that binds Arp2/3 complex to stimulate filament debranching and inhibit actin nucleation. Curr. Biol. 20:1-7. [abstract]
Balcer, H.I., K. Daugherty-Clarke, and B.L. Goode (2010). Mapping surfaces on the p40/ARPC1 subunit of Arp2/3 complex required for actin nucleation and in vivo function. J. Biol. Chem. 285:8481-91. [abstract]
Chaudhry, F., L. Talarico, K. Little, O. Quintero-Monzon, and B.L. Goode (2010). A central role for the WH2 domain of Srv2/CAP in recharging actin monomers to drive actin turnover in vitro and in vivo. Cytoskel. 67:120-33. [abstract]
View Complete Publication List on PubMed: Bruce Goode
Last review: August 9, 2011