Alternative splicing of clathrin heavy chain contributes to the switch from coated pits to plaques

Moulay, Gilles; Lainé, Jeanne; Lemaître, Mégane; Nakamori, Mikihito; Nishino, Ichizo; Caillol, Ghislaine; Mamchaoui, Kamel; Julien, Laura; Dingli, Florent; Loew, Damarys; Bitoun, Marc; Leterrier, Christophe; Furling, Denis; Vassilopoulos, Stéphane

doi:10.1083/jcb.201912061

Cited by 37 publications

(46 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To estimate the curvature of the CCSs, the mean CCS ROI was divided by the average grey value of the PM (as determined by the mean grey value of 4 PM ROIs in each corner of each image). From these two values, the morphology of the CCS could be determined by using thresholds and divided into categories, as described by Moulay et al ., ( 21 ). We used an area threshold of 8500 mm 2 (which is derived from a diameter of 105 nm) to determine if the CCS was small or large, and a curvature value of 1.25 (determined by measuring the mean grey value of the large CCSs observed in TPC disruption conditions) to determine if the CCS was round or flat.…”

Section: Methodsmentioning

confidence: 99%

“…To quantify the effect of TPC disruption, we determined the shape of the clathrin structures by measuring the area and average intensity (a proxy for CCV curvature (Fig. S4A) ( 21 )) of each clathrin structure visualized and classified these shapes into 4 categories; ‘small and round’ (the fully invaginated CCVs), ‘small and flat’ (where curvature generation had failed), ‘large and round’, and ‘large and flat’ (clathrin plaques) (Fig. 3C).…”

Section: Main Textmentioning

confidence: 99%

“…CC-BY-NC 4.0 International license made available under a (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is The copyright holder for this preprint this version posted April 27, 2021. ; https://doi.org/10.1101/2021.04.26.441441 doi: bioRxiv preprint morphology of the CCS could be determined by using thresholds and divided into categories, as described by Moulay et al, (21). We used an area threshold of 8500 mm (which is derived from a diameter of 105 nm) to determine if the CCS was small or large, and a curvature value of 1.25 (determined by measuring the mean grey value of the large CCSs observed in TPC disruption conditions) to determine if the CCS was round or flat.…”

Section: Super-resolution Localization Of Eapsmentioning

confidence: 99%

See 2 more Smart Citations

The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis

Johnson

Dahhan

Gnyliukh

et al. 2021

Preprint

View full text Add to dashboard Cite

Clathrin-mediated endocytosis in plants is an essential process but the underlying mechanisms are poorly understood, not least because of the extreme intracellular turgor pressure acting against the formation of endocytic vesicles. In contrast to other models, plant endocytosis is independent of actin, indicating a mechanistically distinct solution. Here, by using biochemical and advanced microscopy approaches, we show that the plant-specific TPLATE complex acts outside of endocytic vesicles as a mediator of membrane bending. Cells with disrupted TPLATE fail to generate spherical vesicles, and in vitro biophysical assays identified protein domains with membrane bending capability. These results redefine the role of the TPLATE complex as a key component of the evolutionarily distinct mechanism mediating membrane bending against high turgor pressure to drive endocytosis in plant cells.One Sentence SummaryWhile plant CME is actin independent, we identify that the evolutionarily ancient octameric TPLATE complex mediates membrane bending against high turgor pressure in plant clathrin-mediated endocytosis.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Main Textmentioning

confidence: 99%

Section: Super-resolution Localization Of Eapsmentioning

confidence: 99%

See 1 more Smart Citation

The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis

Johnson

Dahhan

Gnyliukh

et al. 2021

Preprint

View full text Add to dashboard Cite

show abstract

“…To estimate the curvature of the CCSs, the mean CCS ROI was divided by the average gray value of the PM (as determined by the mean gray value of 4 PM ROIs in each corner of each image). From these two values, the morphology of the CCS could be determined by using thresholds to divide the CCSs into categories as described by Moulay et al (24). We used an area threshold of 8,500 mm 2 (which is derived from a diameter of 105 nm) to determine if the CCS was small or large and a curvature value of 1.25 (determined by measuring the mean gray value of the large CCSs observed in TPC disruption conditions) to determine if the CCS was round or flat.…”

Section: Methodsmentioning

confidence: 99%

The TPLATE complex mediates membrane bending during plant clathrin–mediated endocytosis

Johnson

Dahhan

Gnyliukh

et al. 2021

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

View full text Add to dashboard Cite

Clathrin-mediated endocytosis is the major route of entry of cargos into cells and thus underpins many physiological processes. During endocytosis, an area of flat membrane is remodeled by proteins to create a spherical vesicle against intracellular forces. The protein machinery which mediates this membrane bending in plants is unknown. However, it is known that plant endocytosis is actin independent, thus indicating that plants utilize a unique mechanism to mediate membrane bending against high-turgor pressure compared to other model systems. Here, we investigate the TPLATE complex, a plant-specific endocytosis protein complex. It has been thought to function as a classical adaptor functioning underneath the clathrin coat. However, by using biochemical and advanced live microscopy approaches, we found that TPLATE is peripherally associated with clathrin-coated vesicles and localizes at the rim of endocytosis events. As this localization is more fitting to the protein machinery involved in membrane bending during endocytosis, we examined cells in which the TPLATE complex was disrupted and found that the clathrin structures present as flat patches. This suggests a requirement of the TPLATE complex for membrane bending during plant clathrin–mediated endocytosis. Next, we used in vitro biophysical assays to confirm that the TPLATE complex possesses protein domains with intrinsic membrane remodeling activity. These results redefine the role of the TPLATE complex and implicate it as a key component of the evolutionarily distinct plant endocytosis mechanism, which mediates endocytic membrane bending against the high-turgor pressure in plant cells.

show abstract

“…In the absence of SMAP-1, we found that the remaining CHC-1-labeled structure no longer colocalized with APG-1 ( Figure 5C-C′ ). In addition to biosynthetic sorting, the clathrin coat is known to mediate the formation of endocytic clathrin-coated vesicles ( Chen and Schmid, 2020 ; Moulay et al, 2020 ). Also, clathrin has been reported to function as a component of the retrograde transport machinery on the surface of the endosome ( Saint-Pol et al, 2004 ; Shi et al, 2009 ).…”

Section: Resultsmentioning

confidence: 99%

AP-1 Recruits SMAP-1/SMAPs to the trans-Golgi Network to Promote Sorting in Polarized Epithelia

Wang

Yao

Zhang

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Coordinated AP-1 and clathrin coat assembly mediate secretory sorting on the trans-Golgi network (TGN) during conventional secretion. Here we found that SMAP-1/SMAPs deficiency caused the apical protein ERM-1 to accumulate on the basolateral side of the TGN. In contrast, the basolateral protein SLCF-1 appeared abnormally on the apical membrane. SMAP-1 colocalized with AP-1 on the TGN. The integrity of AP-1 is required for the subcellular presence of SMAP-1. Moreover, we found that the loss of SMAP-1 reduced clathrin-positive structures in the cytosol, suggesting that SMAP-1 has a regulatory role in clathrin assembly on the TGN. Functional experiments showed that overexpressing clathrin effectively alleviated exocytic defects due to the lack of SMAP-1, corroborating the role of SMAP-1 in promoting the assembly of clathrin on the TGN. Together, our results suggested that the AP-1 complex regulates the TGN localization of SMAP-1, promoting clathrin assembly to ensure polarized conventional secretion in C. elegans intestinal epithelia.

show abstract

Alternative splicing of clathrin heavy chain contributes to the switch from coated pits to plaques

Cited by 37 publications

References 60 publications

The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis

The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis

The TPLATE complex mediates membrane bending during plant clathrin–mediated endocytosis

AP-1 Recruits SMAP-1/SMAPs to the trans-Golgi Network to Promote Sorting in Polarized Epithelia

Contact Info

Product

Resources

About