Bovine brain clathrin light chains impede heavy chain assembly in vitro

Ungewickell, Ernst; Ungewickell, Huberta

doi:10.1016/s0021-9258(18)98957-x

Cited by 69 publications

(15 citation statements)

References 35 publications

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“…One possibility is that Hip1R only partially competes and/or sequesters clathrin LC from coated pits and that the remaining LC can maintain clathrin function. It is interesting to note that clathrin LCs are thought to inhibit HC assembly, so that lattice formation does not spontaneously occur under physiological conditions (Ungewickell and Ungewickell, 1991;Ybe et al, 1998). Since Hip1R binding to truncated clathrin cages depends on LC, Hip1R might promote clathrin assembly via interactions with LC.…”

Section: Discussionmentioning

confidence: 99%

The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro

Engqvist-Goldstein

Warren

Kessels

et al. 2001

The Journal of Cell Biology

242

224

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Huntingtin-interacting protein 1 related (Hip1R) is a novel component of clathrin-coated pits and vesicles and is a mammalian homologue of Sla2p, an actin-binding protein important for both actin organization and endocytosis in yeast. Here, we demonstrate that Hip1R binds via its putative central coiled-coil domain to clathrin, and provide evidence that Hip1R and clathrin are associated in vivo at sites of endocytosis. First, real-time analysis of Hip1R–YFP and DsRed–clathrin light chain (LC) in live cells revealed that these proteins show almost identical temporal and spatial regulation at the cell cortex. Second, at the ultrastructure level, immunogold labeling of ‘unroofed’ cells showed that Hip1R localizes to clathrin-coated pits. Third, overexpression of Hip1R affected the subcellular distribution of clathrin LC. Consistent with a functional role for Hip1R in endocytosis, we also demonstrated that it promotes clathrin cage assembly in vitro. Finally, we showed that Hip1R is a rod-shaped apparent dimer with globular heads at either end, and that it can assemble clathrin-coated vesicles and F-actin into higher order structures. In total, Hip1R's properties suggest an early endocytic function at the interface between clathrin, F-actin, and lipids.

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Section: Discussionmentioning

confidence: 99%

The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro

Engqvist-Goldstein

Warren

Kessels

et al. 2001

The Journal of Cell Biology

242

224

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“…In vitro , clathrin triskelia assemble into spherical cages 13 . Clathrin light chain inhibits this assembly 24, 25 . Light chain binds to heavy chain through multiple interactions 16 .…”

Section: Introductionmentioning

confidence: 99%

A conformational switch in clathrin light chain regulates lattice structure and endocytosis at the plasma membrane of mammalian cells

Obashi

Sochacki

Strub

et al. 2022

Preprint

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The conformations of endocytic proteins and their interactions are key regulators of clathrin-mediated endocytosis. Three clathrin light chains (CLC), along with three clathrin heavy chains, assemble to form single clathrin triskelia that link into a geometric lattice that curves to drive endocytosis. Conformational changes in CLC have been shown to regulate triskelia assembly in solution, yet the nature of these structural changes, and their effects on lattice growth, curvature, and endocytosis in cells are unclear. Here, we develop a correlative fluorescence resonance energy transfer (FRET) and platinum replica electron microscopy method, named FRET-CLEM. With FRET-CLEM, we measure conformational changes in proteins at thousands of individual morphologically distinct clathrin-coated structures across cell membranes. We find that the N-terminus of CLC moves away from the plasma membrane and triskelia vertex as lattices curve. Preventing this conformational switch with acute chemical tools inside cells increased clathrin structure sizes and inhibited endocytosis. Therefore, a specific conformational switch in CLC regulates lattice curvature and endocytosis in mammalian cells.

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“…A clue to the function of the clathrin light chains came from the observation that light-chain-free triskelia showed a greater propensity for self-association than those with attached light chains [10,13,14]. The light chains might thus act to prevent premature or futile assembly of triskelia in the cytosol.…”

mentioning

confidence: 99%