Structural Basis of Actin Filament Nucleation by Tandem W Domains

Overview

Journal Cell Rep

Publisher Cell Press

Specialties Biology
Cell Biology
Molecular Biology

Date 2013 Jun 4

PMID 23727244

Citations 14

Authors

Xiaorui Chen

Fengyun Ni

Xia Tian

Elena Kondrashkina

Qinghua Wang

Jianpeng Ma

Affiliations

Soon will be listed here.

Abstract

Spontaneous nucleation of actin is very inefficient in cells. To overcome this barrier, cells have evolved a set of actin filament nucleators to promote rapid nucleation and polymerization in response to specific stimuli. However, the molecular mechanism of actin nucleation remains poorly understood. This is hindered largely by the fact that actin nucleus, once formed, rapidly polymerizes into filament, thus making it impossible to capture stable multisubunit actin nucleus. Here, we report an effective double-mutant strategy to stabilize actin nucleus by preventing further polymerization. Employing this strategy, we solved the crystal structure of AMPPNP-actin in complex with the first two tandem W domains of Cordon-bleu (Cobl), a potent actin filament nucleator. Further sequence comparison and functional studies suggest that the nucleation mechanism of Cobl is probably shared by the p53 cofactor JMY, but not Spire. Moreover, the double-mutant strategy opens the way for atomic mechanistic study of actin nucleation and polymerization.

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References

Rould M, Wan Q, Joel P, Lowey S, Trybus K . Crystal structures of expressed non-polymerizable monomeric actin in the ADP and ATP states. J Biol Chem. 2006; 281(42):31909-19. DOI: 10.1074/jbc.M601973200. View

Sitar T, Gallinger J, Ducka A, Ikonen T, Wohlhoefler M, Schmoller K . Molecular architecture of the Spire-actin nucleus and its implication for actin filament assembly. Proc Natl Acad Sci U S A. 2011; 108(49):19575-80. PMC: 3241762. DOI: 10.1073/pnas.1115465108. View

Husson C, Cantrelle F, Roblin P, Didry D, Le K, Perez J . Multifunctionality of the beta-thymosin/WH2 module: G-actin sequestration, actin filament growth, nucleation, and severing. Ann N Y Acad Sci. 2010; 1194:44-52. DOI: 10.1111/j.1749-6632.2010.05473.x. View

Irobi E, Aguda A, Larsson M, Guerin C, Yin H, Burtnick L . Structural basis of actin sequestration by thymosin-beta4: implications for WH2 proteins. EMBO J. 2004; 23(18):3599-608. PMC: 517612. DOI: 10.1038/sj.emboj.7600372. View

Otomo T, Tomchick D, Otomo C, Panchal S, Machius M, Rosen M . Structural basis of actin filament nucleation and processive capping by a formin homology 2 domain. Nature. 2005; 433(7025):488-94. DOI: 10.1038/nature03251. View

Husson C, Renault L, Didry D, Pantaloni D, Carlier M . Cordon-Bleu uses WH2 domains as multifunctional dynamizers of actin filament assembly. Mol Cell. 2011; 43(3):464-77. DOI: 10.1016/j.molcel.2011.07.010. View

Campellone K, Welch M . A nucleator arms race: cellular control of actin assembly. Nat Rev Mol Cell Biol. 2010; 11(4):237-51. PMC: 2929822. DOI: 10.1038/nrm2867. View

Carroll E, Gerrelli D, Gasca S, Berg E, Beier D, Copp A . Cordon-bleu is a conserved gene involved in neural tube formation. Dev Biol. 2003; 262(1):16-31. DOI: 10.1016/s0012-1606(03)00323-3. View

Otterbein L, Graceffa P, Dominguez R . The crystal structure of uncomplexed actin in the ADP state. Science. 2001; 293(5530):708-11. DOI: 10.1126/science.1059700. View

10.

. The CCP4 suite: programs for protein crystallography. Acta Crystallogr D Biol Crystallogr. 1994; 50(Pt 5):760-3. DOI: 10.1107/S0907444994003112. View

11.

Spudich J, Watt S . The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971; 246(15):4866-71. View

12.

Oda T, Iwasa M, Aihara T, Maeda Y, Narita A . The nature of the globular- to fibrous-actin transition. Nature. 2009; 457(7228):441-5. DOI: 10.1038/nature07685. View

13.

Iwasa M, Maeda K, Narita A, Maeda Y, Oda T . Dual roles of Gln137 of actin revealed by recombinant human cardiac muscle alpha-actin mutants. J Biol Chem. 2008; 283(30):21045-53. PMC: 3258932. DOI: 10.1074/jbc.M800570200. View

14.

Rebowski G, Boczkowska M, Hayes D, Guo L, Irving T, Dominguez R . X-ray scattering study of actin polymerization nuclei assembled by tandem W domains. Proc Natl Acad Sci U S A. 2008; 105(31):10785-90. PMC: 2504828. DOI: 10.1073/pnas.0801650105. View

15.

Sablin E, Dawson J, VanLoock M, Spudich J, Egelman E, Fletterick R . How does ATP hydrolysis control actin's associations?. Proc Natl Acad Sci U S A. 2002; 99(17):10945-7. PMC: 123189. DOI: 10.1073/pnas.152329899. View

16.

Zuchero J, Coutts A, Quinlan M, La Thangue N, Mullins R . p53-cofactor JMY is a multifunctional actin nucleation factor. Nat Cell Biol. 2009; 11(4):451-9. PMC: 2763628. DOI: 10.1038/ncb1852. View

17.

Kabsch W, Mannherz H, Suck D, Pai E, Holmes K . Atomic structure of the actin:DNase I complex. Nature. 1990; 347(6288):37-44. DOI: 10.1038/347037a0. View

18.

Firat-Karalar E, Hsiue P, Welch M . The actin nucleation factor JMY is a negative regulator of neuritogenesis. Mol Biol Cell. 2011; 22(23):4563-74. PMC: 3226475. DOI: 10.1091/mbc.E11-06-0585. View

19.

Ravanelli A, Klingensmith J . The actin nucleator Cordon-bleu is required for development of motile cilia in zebrafish. Dev Biol. 2010; 350(1):101-11. PMC: 3022090. DOI: 10.1016/j.ydbio.2010.11.023. View

20.

Chen C, Sawaya M, Phillips M, Reisler E, Quinlan M . Multiple forms of Spire-actin complexes and their functional consequences. J Biol Chem. 2012; 287(13):10684-10692. PMC: 3323035. DOI: 10.1074/jbc.M111.317792. View