Real-Time Visualization of Dynamin-Catalyzed Membrane Fission and Vesicle Release

Pucadyil, Thomas J.; Schmid, Sandra L.

doi:10.1016/j.cell.2008.11.020

Cited by 258 publications

(312 citation statements)

References 34 publications

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“…A striking feature of many BAR/F-BAR domain proteins is the concomitant presence of at least one SH3 domain (3). In the case of endocytic BAR/F-BAR domain proteins including syndapin, endophilin, and amphiphysin, the SH3 domain acts to recruit proline-rich ligands, most notably the large membranedeforming and -scissioning GTPase dynamin, a key factor in endocytic membrane fission (10,11,30,31).…”

Section: Association Of Syndapin 1 With Dynamin 1 or A Dynamin 1-derimentioning

confidence: 99%

Molecular basis for SH3 domain regulation of F-BAR–mediated membrane deformation

Rao

Vahedi-Faridi

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

140

182

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Members of the Bin/amphiphysin/Rvs (BAR) domain protein superfamily are involved in membrane remodeling in various cellular pathways ranging from endocytic vesicle and T-tubule formation to cell migration and neuromorphogenesis. Membrane curvature induction and stabilization are encoded within the BAR or Fer-CIP4 homology-BAR (F-BAR) domains, α-helical coiled coils that dimerize into membrane-binding modules. BAR/F-BAR domain proteins often contain an SH3 domain, which recruits binding partners such as the oligomeric membrane-fissioning GTPase dynamin. How precisely BAR/F-BAR domain-mediated membrane deformation is regulated at the cellular level is unknown. Here we present the crystal structures of full-length syndapin 1 and its F-BAR domain. Our data show that syndapin 1 F-BAR-mediated membrane deformation is subject to autoinhibition by its SH3 domain. Release from the clamped conformation is driven by association of syndapin 1 SH3 with the proline-rich domain of dynamin 1, thereby unlocking its potent membrane-bending activity. We hypothesize that this mechanism might be commonly used to regulate BAR/F-BAR domain-induced membrane deformation and to potentially couple this process to dynamin-mediated fission. Our data thus suggest a structure-based model for SH3-mediated regulation of BAR/F-BAR domain function.

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Section: Association Of Syndapin 1 With Dynamin 1 or A Dynamin 1-derimentioning

confidence: 99%

Molecular basis for SH3 domain regulation of F-BAR–mediated membrane deformation

Rao

Vahedi-Faridi

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

140

182

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“…Growing CCPs acquire cargo and invaginate via clathrin polymerization [4] and the coordinated action of curvature-inducing/sensing BAR [5] and F-BAR domain proteins [6,7], ENTH domain proteins [8], and possibly actin [9][10][11]. The neck of the deeply invaginated CCP is severed in a mechanism involving the large GTPase dynamin [12,13], and possibly a phosphoinositide (PI) phosphatase [14], to release a clathrin-coated vesicle (CCV), which uncoats through the action of GAK/auxilin [15,16].…”

Section: Introductionmentioning

confidence: 99%

A High Precision Survey of the Molecular Dynamics of Mammalian Clathrin-Mediated Endocytosis

Perrais²,

2011

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Dual colour total internal reflection fluorescence microscopy is a powerful tool for decoding the molecular dynamics of clathrin-mediated endocytosis (CME). Typically, the recruitment of a fluorescent protein-tagged endocytic protein was referenced to the disappearance of spot-like clathrin-coated structure (CCS), but the precision of spot-like CCS disappearance as a marker for canonical CME remained unknown. Here we have used an imaging assay based on total internal reflection fluorescence microscopy to detect scission events with a resolution of ,2 s. We found that scission events engulfed comparable amounts of transferrin receptor cargo at CCSs of different sizes and CCS did not always disappear following scission. We measured the recruitment dynamics of 34 types of endocytic protein to scission events: Abp1, ACK1, amphiphysin1, APPL1, Arp3, BIN1, CALM, CIP4, clathrin light chain (Clc), cofilin, coronin1B, cortactin, dynamin1/ 2, endophilin2, Eps15, Eps8, epsin2, FBP17, FCHo1/2, GAK, Hip1R, lifeAct, mu2 subunit of the AP2 complex, myosin1E, myosin6, NECAP, N-WASP, OCRL1, Rab5, SNX9, synaptojanin2b1, and syndapin2. For each protein we aligned ,1,000 recruitment profiles to their respective scission events and constructed characteristic ''recruitment signatures'' that were grouped, as for yeast, to reveal the modular organization of mammalian CME. A detailed analysis revealed the unanticipated recruitment dynamics of SNX9, FBP17, and CIP4 and showed that the same set of proteins was recruited, in the same order, to scission events at CCSs of different sizes and lifetimes. Collectively these data reveal the fine-grained temporal structure of CME and suggest a simplified canonical model of mammalian CME in which the same core mechanism of CME, involving actin, operates at CCSs of diverse sizes and lifetimes.

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“…Dynamin is a large GTPase that has been shown to polymerize into a helical collar at the neck of endocytic buds (3), where it subsequently plays a key role in the formation of endocytic vesicles through fission (4)(5)(6)(7)(8). This function is fundamental, as the knockout of the dynamin neuronal isoform leads to striking defects in synapse organization and results in a strong dysfunction of neuronal activity (9).…”

mentioning

confidence: 99%

“…This function is fundamental, as the knockout of the dynamin neuronal isoform leads to striking defects in synapse organization and results in a strong dysfunction of neuronal activity (9). The recruitment of dynamin to endocytic buds is thought to depend on the local synthesis of phosphatidylinositol (4,5)bisphosphate (PIP 2 ), as dynamin has a PIP 2 binding pleckstrin homology (PH) domain (10). Dynamin is recruited late in clathrin-coated vesicle formation, as seen by total internal reflection fluorescence (TIRF) microscopy (11)(12)(13).…”

mentioning

confidence: 99%

Membrane curvature controls dynamin polymerization

Roux

Koster

Lenz

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

271

263

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The generation of membrane curvature in intracellular traffic involves many proteins that can curve lipid bilayers. Among these, dynamin-like proteins were shown to deform membranes into tubules, and thus far are the only proteins known to mechanically drive membrane fission. Because dynamin forms a helical coat circling a membrane tubule, its polymerization is thought to be responsible for this membrane deformation. Here we show that the force generated by dynamin polymerization, 18 pN, is sufficient to deform membranes yet can still be counteracted by high membrane tension. Importantly, we observe that at low dynamin concentration, polymer nucleation strongly depends on membrane curvature. This suggests that dynamin may be precisely recruited to membrane buds' necks because of their high curvature. To understand this curvature dependence, we developed a theory based on the competition between dynamin polymerization and membrane mechanical deformation. This curvature control of dynamin polymerization is predicted for a specific range of concentrations (∼0.1-10 μM), which corresponds to our measurements. More generally, we expect that any protein that binds or self-assembles onto membranes in a curvature-coupled way should behave in a qualitatively similar manner, but with its own specific range of concentration.force | nucleation | fission | endocytosis | dynamin-like proteins M embrane remodeling is an essential task of proteins involved in membrane traffic (1, 2). Dynamin is a large GTPase that has been shown to polymerize into a helical collar at the neck of endocytic buds (3), where it subsequently plays a key role in the formation of endocytic vesicles through fission (4-8). This function is fundamental, as the knockout of the dynamin neuronal isoform leads to striking defects in synapse organization and results in a strong dysfunction of neuronal activity (9). The recruitment of dynamin to endocytic buds is thought to depend on the local synthesis of phosphatidylinositol(4,5)bisphosphate (PIP 2 ), as dynamin has a PIP 2 binding pleckstrin homology (PH) domain (10). Dynamin is recruited late in clathrin-coated vesicle formation, as seen by total internal reflection fluorescence (TIRF) microscopy (11-13). Because PIP 2 is also responsible for the binding of clathrin coats, it is expected to be present at the clathrin bud from the beginning of its formation. Thus, another explanation was suggested: Proteins that interact with dynamin and possess a curvature-sensing Bin-Amphiphysin-Rvs (BAR) domain (such as endophilin and amphiphysin) were proposed to sense the high curvature of the neck and recruit dynamin (14). Because the curvature of the neck is increasing during clathrin-coated vesicle formation to finally reach the range needed for BAR recruitment, this could explain the arrival of dynamin at the very late stage of clathrin-coated vesicle formation. However, in solution, dynamin spontaneously associates into helices and rings (15) with an internal radius of ≈10 nm. Dynamin can also polymerize around pre...

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Real-Time Visualization of Dynamin-Catalyzed Membrane Fission and Vesicle Release

Cited by 258 publications

References 34 publications

Molecular basis for SH3 domain regulation of F-BAR–mediated membrane deformation

Molecular basis for SH3 domain regulation of F-BAR–mediated membrane deformation

A High Precision Survey of the Molecular Dynamics of Mammalian Clathrin-Mediated Endocytosis

Membrane curvature controls dynamin polymerization

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