In eukaryotes, clathrin-coated vesicles (CCVs) facilitate the internalization of material from the cell surface as well as the movement of cargo in post-Golgi trafficking pathways. This diversity of functions is partially provided by multiple monomeric and multimeric clathrin adaptor complexes that provide compartment and cargo selectivity. The adaptor-protein assembly polypeptide-1 (AP-1) complex operates as part of the secretory pathway at the trans-Golgi network (TGN), while the AP-2 complex and the TPLATE complex jointly operate at the plasma membrane to execute clathrin-mediated endocytosis. Key to our further understanding of clathrin-mediated trafficking in plants will be the comprehensive identification and characterization of the network of evolutionarily conserved and plant-specific core and accessory machinery involved in the formation and targeting of CCVs. To facilitate these studies, we have analyzed the proteome of enriched TGN/early endosome-derived and endocytic CCVs isolated from dividing and expanding suspension-cultured Arabidopsis (Arabidopsis thaliana) cells. Tandem mass spectrometry analysis results were validated by differential chemical labeling experiments to identify proteins co-enriching with CCVs. Proteins enriched in CCVs included previously characterized CCV components and cargos such as the vacuolar sorting receptors in addition to conserved and plant-specific components whose function in clathrin-mediated trafficking has not been previously defined. Notably, inmore »
- Award ID(s):
- 2114603
- Publication Date:
- NSF-PAR ID:
- 10337441
- Journal Name:
- The Plant Cell
- ISSN:
- 1532-298X
- Sponsoring Org:
- National Science Foundation
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Abstract -
Endocytosis, secretion, and endosomal trafficking are key cellular processes that control the composition of the plasma membrane. Through the coordination of these trafficking pathways, cells can adjust the composition, localization, and turnover of proteins and lipids in response to developmental or environmental cues. Upon being incorporated into vesicles and internalized through endocytosis, plant plasma membrane proteins are delivered to the trans‐Golgi network (TGN). At the TGN, plasma membrane proteins are recycled back to the plasma membrane or transferred to multivesicular endosomes (MVEs), where they are further sorted into intralumenal vesicles for degradation in the vacuole. Both types of plant endosomes, TGN and MVEs, act as sorting organelles for multiple endocytic, recycling, and secretory pathways. Molecular assemblies such as retromer, ESCRT (endosomal sorting complex required for transport) machinery, small GTPases, adaptor proteins, and SNAREs associate with specific domains of endosomal membranes to mediate different sorting and membrane‐budding events. In this review, we discuss the mechanisms underlying the recognition and sorting of proteins at endosomes, membrane remodeling and budding, and their implications for cellular trafficking and physiological responses in plants.
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Key points Association of plasma membrane BKCachannels with BK‐β subunits shapes their biophysical properties and physiological roles; however, functional modulation of the mitochondrial BKCachannel (mitoBKCa) by BK‐β subunits is not established.
MitoBKCa‐α and the regulatory BK‐β1 subunit associate in mouse cardiac mitochondria.
A large fraction of mitoBKCadisplay properties similar to that of plasma membrane BKCawhen associated with BK‐β1 (left‐shifted voltage dependence of activation,
V 1/2 = −55 mV, 12 µm matrix Ca2+).In BK‐β1 knockout mice, cardiac mitoBKCadisplayed a low
P oand a depolarizedV 1/2of activation (+47 mV at 12 µm matrix Ca2+)Co‐expression of BKCawith the BK‐β1 subunit in HeLa cells doubled the density of BKCain mitochondria.
The present study supports the view that the cardiac mitoBKCachannel is functionally modulated by the BK‐β1 subunit; proper targeting and activation of mitoBKCashapes mitochondrial Ca2+handling.
Abstract Association of the plasma membrane BKCachannel with auxiliary BK‐β1–4 subunits profoundly affects the regulatory mechanisms and physiological processes in which this channel participates. However, functional association of mitochondrial BK (mitoBKCa) with regulatory subunits is unknown. We report that mitoBKCafunctionally associates with its regulatory subunit BK‐β1 in adult rodent cardiomyocytes. Cardiac mitoBKCais a calcium‐ and voltage‐activated channel that is sensitive to paxilline with a large conductance for K+of 300 pS. Additionally, mitoBKCadisplays a high open probability (
P o) and voltage half‐activation (V 1/2 = −55 mV,n = 7) resembling that of plasma membrane BKCawhenmore » -
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