Spatial control of coated-pit dynamics in living cells.
Here we visualize new aspects of the dynamics of endocytotic clathrin-coated pits and vesicles in mammalian cells by using a fusion protein consisting of green fluorescent protein and clathrin light chain a. Clathrin-coated pits invaginating from the plasma membrane show definite, but highly limited, mobility within the membrane that is relaxed upon treatment with latrunculin B, an inhibitor of actin assembly, indicating that an actin-based framework may be involved in the mobility of these pits. Transient, motile coated vesicles that originate from coated pits can be detected, with multiple vesicles occasionally appearing to emanate from a single pit. Despite their seemingly random distribution, coated pits tend to form repeatedly at defined sites while excluding other regions. This spatial regulation of coated-pit assembly and function is attributable to the attachment of the coated pits to the membrane skeleton.
Gaidarov
I
I
Kimmel Cancer Institute, Philadelphia, Pennsylvania, USA.
Santini
F
F
Warren
R A
RA
Keen
J H
JH
D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D019556:COS Cells; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D002966:Clathrin / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D004705:Endocytosis; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D008387:Marine Toxins / Q000494:pharmacology; D051379:Mice; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines
D019086:Bridged Bicyclo Compounds, Heterocyclic; D002966:Clathrin; D008164:Luminescent Proteins; D008387:Marine Toxins; D011993:Recombinant Fusion Proteins; D013844:Thiazoles; D053778:Thiazolidines; D049452:Green Fluorescent Proteins; C037068:latrunculin B