GTPase Cycle of Dynamin Is Coupled to Membrane Squeeze and Release, Leading to Spontaneous Fission

Bashkirov, Pavel V.; Akimov, Sergey A.; Evseev, A. I.; Schmid, Sandra L.; Zimmerberg, Joshua; Frolov, Vadim A.

doi:10.1016/j.cell.2008.11.028

Cited by 276 publications

(401 citation statements)

References 42 publications

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“…Critically, scission was found to be far more efficient if the membrane tubule was placed under tension, most likely because this eliminated the dissipation of dynamin-induced torque by membrane flow and instead favored membrane shearing (Morlot and Roux 2013). Most likely, a transient hemifused state leads to scission because the scission reaction is not leaky, as determined using elegant conductance measurements (Bashkirov et al 2008). Overall, these results mesh well with recent studies publishing the crystal structure of dynamin, which provided detailed insight into the GTP-hydrolysis constriction mechanism of oligomerized dynamin (Chappie et al 2011;Faelber et al 2011Faelber et al , 2012Ford et al 2011).…”

Section: Membrane Scissionmentioning

confidence: 99%

Endocytic Accessory Factors and Regulation of Clathrin-Mediated Endocytosis

Merrifield

Kaksonen

2014

Cold Spring Harbor Perspectives in Biology

114

111

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Section: Membrane Scissionmentioning

confidence: 99%

Endocytic Accessory Factors and Regulation of Clathrin-Mediated Endocytosis

Merrifield

Kaksonen

2014

Cold Spring Harbor Perspectives in Biology

114

111

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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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“…4a). The identified oligomerization sites in interface-3 include two loop regions (L1, L2) which, together with the proposed interaction in L4, might allow some flexibility in the degree of rotation, concomitant with varying ring diameters as observed for oligomerised dynamin and MxA 23,29 (Fig. 4c).…”

mentioning

confidence: 99%

Structural basis of oligomerization in the stalk region of dynamin-like MxA

Gao

Malsburg

Paeschke

et al. 2010

Nature

225

428

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GTPase is a key mediator of cell-autonomous innate immunityHis-tagged full-length human MxA (Fig. 1a) was recombinantly expressed in bacteria and purified to homogeneity (Methods, Supp. Fig. 1). In crystallization trials, small needle-shaped protein crystals were obtained which represented proteolytic cleavage products of the MD and GED (Supp. Fig. 2). We solved the phase problem by a single anomalous dispersion protocol and could build and refine a model containing two molecules in the asymmetric unit (Methods, Supp. Table 1 and 2). Each monomer spans nearly the complete MD and the amino(N-)-terminal part of the GED (amino acids 366-633) which together fold into an elongated anti-parallel fourhelical bundle where the MD contributes three helices and the GED one (Fig. 1b, Supp. Fig. 3). This segment corresponds to the stalk region of dynamin 7 , and we refer to it as stalk of MxA. The first visible amino acid, Glu366, is 15 amino acids downstream of the last visible residue of the corresponding G-domain structure in rat dynamin (Supp. Fig. 3) 8 . It marks the start of helix α1 in the MxA stalk which is divided in α1 N and α1 C by a 10 amino acid long loop, L1, introducing a 30° kink. A putative loop L2 (amino acids 438-447) opposite of the deduced position of the G-domain is not visible in our structure. L2 was previously demonstrated to be the target of a functionally neutralising monoclonal antibody 9,10 . Helix α2 runs anti-parallel to α1 back to the G-domain. It ends in a short loop L3 and is followed by helix α3 that extends in parallel to α1. The 40 amino acid long loop L4 (residues 532-572) is at the equivalent sequence position as the PH domain of dynamin (Fig. 1a, Supp. Fig. 3) and is absent in our model. L4 is predicted to be unstructured and was previously shown to be proteinase K sensitive 11 .At the C-terminus, the GED supplies 44 residues to helix α4 which proceeds in parallel to helix α2 back to the G-domain. It is followed by a short helix α5 which directs the polypeptide chain towards the N-terminus of the MD. The carboxy(C-)-terminal 30 highly conserved residues of the GED known to be involved in antiviral specificity 12 are missing in our model. In dynamin, the corresponding residues were shown to directly interact with the G-domain 13 . The stalk of MxA is divergent from the corresponding structures of other dynamin superfamily members, such as GBP1 14 , EHD2 15 and BDLP 16 although some features are shared (Supp. Fig. 4). 4 In the crystal lattice, each MxA stalk is assembled in a criss-cross pattern resulting in a linear oligomer, where each stalk contributes three distinct interfaces (Fig. 1c). Such an arrangement of the stalks is plausible for the Mx oligomer since all G-domains would be located at one side of the oligomer whereas the putative substrate-binding site in L2 and L4 would be located at the opposite side (Fig. 1b, c).The hydrophobic interface-1 covering 1300 Å 2 is conserved among Mx proteins and dynamins and has a two-fold symmetry between the associating monomers (Fig....

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GTPase Cycle of Dynamin Is Coupled to Membrane Squeeze and Release, Leading to Spontaneous Fission

Cited by 276 publications

References 42 publications

Endocytic Accessory Factors and Regulation of Clathrin-Mediated Endocytosis

Endocytic Accessory Factors and Regulation of Clathrin-Mediated Endocytosis

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

Structural basis of oligomerization in the stalk region of dynamin-like MxA

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