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  • Review Article
  • Published:

Three ways to make a vesicle

Nature Reviews Molecular Cell Biology volume 1, pages 187–198 (2000)Cite this article

Key Points

  • Some steps of membrane transport require the formation of vesicles coated with COPI, COPII or clathrin. The coats probably generate the forces necessary to bend a relatively flat membrane into a vesicle.

  • COPI coats function in transport through the Golgi and in retrograde transport from the Golgi to the endoplasmic reticulum. COPII coats function in transport from the ER to the Golgi. Clathrin functions in receptor-mediated endocytosis at the plasma membrane when associated with AP2 adaptors, and in transport from the trans-Golgi network (TGN) to endosomes when associated with AP1 adaptors.

  • The assembly of COPII coats starts with the activation of the small GTPase Sar1p, followed by the sequential recruitment of Sec24p-Sec23p and then Sec13p-Sec31p. The GTPase activity of Sar1p acts as a timer for coat release.

  • Similarly, the assembly of COPI coats starts with the activation of the small GTPase Arf1, followed by the recruitment of the coatomer, which binds to the cytoplasmic domain of cargo. The GTPase activity of Arf1 acts as a timer for coat release.

  • The formation of clathrin coats is more complicated, as coat assembly, vesicle budding and uncoating are distinguishable steps regulated by different sets of proteins.

  • Adaptor proteins (AP1, AP2 and arrestins) bind to the cytoplasmic signal sequences of cargo molecules. The recruitment of adaptors is only energy dependent at the TGN, and not at the plasma membrane.

  • Many accessory proteins take part in a network of interactions that regulates clathrin-coat formation. In most cases, the precise function of these proteins is not known.

  • Vesicle scission requires GTP hydrolysis by the GTPase dynamin.

  • Vesicle uncoating requires ATP hydrolysis by auxilin-hsc70.

  • The structure of many of the components of the clathrin machinery has been solved, and many of the protein-protein interactions have been analysed at atomic resolution. However, no structural information is available for COPI and COPII coats.

Abstract

Cargo molecules have to be included in carrier vesicles of different forms and sizes to be transported between organelles. During this process, a limited set of proteins, including the coat proteins COPI, COPII and clathrin, carries out a programmed set of sequential interactions that lead to the budding of vesicles. A general model to explain the formation of coated vesicles is starting to emerge but the picture is more complex than we had imagined.

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Figure 1: The major membrane traffic pathways that use carrier vesicles coated with COPI, COPII and clathrin in eukaryotic cells
Figure 2: The key steps in the formation of COPII-coated vesicles.
Figure 3: The key steps in the formation of COPI-coated vesicles.
Figure 4: The key steps in the formation of clathrin-coated vesicles.
Figure 5: Network of protein–protein and protein–lipid interactions involved in clathrin coat formation at the plasma membrane.
Figure 6: Clathrin coats, a collage of medium and high-resolution views.

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    Tomas Kirchhausen

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Computer simulation of how a clathrin coat might assemble or disassemble (if run in reverse). The animation is based on geometrical constraints imposed by the three-dimensional shape of clathrin triskelions determined in solution by electron microscopy 99, the model of a clathrin coat determined by electron cryomicroscopy 93 and the fit of the atomic structure of the clathrin terminal domain solved by x-ray crystallography 82 to the model of the coat 94. Clathrin triskelions have a characteristic pinwheel appearance; each of the three legs contains a heavy chain and a light chain, spanning a total length of 500 Å. A fully assembled coat contains one triskelion centre at each vertex so that every leg spans two adjacent edges. The music is "Prelude in C minor" by Rachmaninoff. The animation will be presented in a group exhibition of the ART-SCIENCE FUSION project during the 5th International Annaberg Conference in Goldegg castle. This is a joint project with the 'Kulturverein Schloss Goldegg'. Schloss Goldegg focus its 2000/2001 projects on ‚ELEMENTAR'. The exhibition will then move to 'Monique Goldstrom Gallery', 560 Broadway, New York, USA

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DATABASE LINKS

Sec23p

Sec24p

Sec13p

Sec31p

Sar1p

Sec12p

Emp24

COPI coatamer

KDEL receptor

ARF1

ARNO3

ARFGAP

LDL receptor

clathrin

AP-1

AP-2

arrestins

synaptotagmin

amphiphysin

epsin

synaptojanin

Eps15

dynamin

Hsc70

endophilin

phospholipase D

EH domain

ENTH domain

Pleckstrin homology domain

ENCYCLOPEDIA OF LIFE SCIENCES

Clathrin-coated vesicles and receptor-mediated endocytosis

Further reading

Accessory factors in clathrin-dependent synaptic vesicle endocytosis

Glossary

ENDOCYTIC PATHWAY

Macromolecules are taken up by invagination of the plasma membrane. They first arrive in early endosomes, then late endosomes, and finally lysosomes, where they are degraded by hydrolases.

SECRETORY PATHWAY

Secretory or membrane proteins are inserted into the endoplasmic reticulum. They are then transported through the Golgi to the trans-Golgi network, where they are sorted to their final destination.

FIBROBLAST

Common cell type found in connective tissue in many parts of the body, which secretes an extracellular matrix rich in collagen and other macromolecules and connects cell layers.

MACROPINOCYTOSIS

Actin-dependent process by which cells engulf large volumes of fluids.

PHAGOCYTOSIS

Actin-dependent process, by which cells engulf external particulate material by extension and fusion of pseudopods around each particle.

CAVEOLA

Flask-shaped invagination at the plasma membrane, possibly involved in the uptake of extracellular materials.

GREEN-FLUORESCENT PROTEIN

Autofluorescent protein originally identified in the jellyfish Aequorea victoria.

MYRISTOYLATION

Covalent attachment of a hydrophobic myristoyl group to the amino-terminal glycine residue of a nascent polypeptide.

J DOMAIN

Approximately 73 amino-acid region found in DnaJ-like heat shock proteins, which catalytically activates proteins of the Hsc70 family.

SH3 DOMAINS

(Src homology region 3 domains.) Protein sequences of about 50 amino acids that recognize and bind sequences rich in proline.

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Kirchhausen, T. Three ways to make a vesicle. Nat Rev Mol Cell Biol 1, 187–198 (2000). https://doi.org/10.1038/35043117

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