Assembly and targeting of adaptin chimeras in transfected cells - PubMed (original) (raw)

Assembly and targeting of adaptin chimeras in transfected cells

M S Robinson. J Cell Biol. 1993 Oct.

Abstract

Adaptors are the components of clathrincoated pits and vesicles that attach the clathrin to the membrane. There are two types of adaptors in the cell: one associated with the plasma membrane and one associated with the TGN. Both adaptors are heterotetramers consisting of two adaptins (alpha and beta for the plasma membrane; gamma and beta' for the TGN), plus two smaller proteins. The COOH-terminal domains of the adaptins form appendages that resemble ears, connected by flexible hinges. Unlike the other adaptor components, the COOH termini of the alpha- and gamma-adaptins show no homology with each other, suggesting that they might provide the signal that directs the adaptors to the appropriate membrane. To test this possibility, the COOH-terminal ears were switched between alpha- and gamma-adaptins and were also deleted. All of the constructs contained the bovine gamma-adaptin hinge, enabling them to be detected with a species-specific antibody against this region when transfected into rat fibroblasts. Immunoprecipitation indicated that the engineered adaptins were still fully capable of assembling into adaptor complexes. Immunofluorescence revealed that in spite of their modified ears, the constructs were still able to be recruited onto the appropriate membrane; however, the ear-minus constructs gave increased cytoplasmic staining, and replacing the gamma-adaptin ear with the alpha-adaptin ear caused a small amount of colocalization with endogenous alpha-adaptin in some cells. Thus, the major targeting determinant appears to reside in the adaptor "head," while the ears may stabilize the association of adaptors with the membrane.

PubMed Disclaimer

Cited by

Assembly and function of AP-3 complexes in cells expressing mutant subunits.
Peden AA, Rudge RE, Lui WW, Robinson MS. Peden AA, et al. J Cell Biol. 2002 Jan 21;156(2):327-36. doi: 10.1083/jcb.200107140. Epub 2002 Jan 21. J Cell Biol. 2002. PMID: 11807095 Free PMC article.
Interaction of the bovine papillomavirus E6 protein with the clathrin adaptor complex AP-1.
Tong X, Boll W, Kirchhausen T, Howley PM. Tong X, et al. J Virol. 1998 Jan;72(1):476-82. doi: 10.1128/JVI.72.1.476-482.1998. J Virol. 1998. PMID: 9420248 Free PMC article.
Transient Fcho1/2⋅Eps15/R⋅AP-2 Nanoclusters Prime the AP-2 Clathrin Adaptor for Cargo Binding.
Ma L, Umasankar PK, Wrobel AG, Lymar A, McCoy AJ, Holkar SS, Jha A, Pradhan-Sundd T, Watkins SC, Owen DJ, Traub LM. Ma L, et al. Dev Cell. 2016 Jun 6;37(5):428-43. doi: 10.1016/j.devcel.2016.05.003. Epub 2016 May 26. Dev Cell. 2016. PMID: 27237791 Free PMC article.
An internally eGFP-tagged α-adaptin is a fully functional and improved fiduciary marker for clathrin-coated pit dynamics.
Mino RE, Chen Z, Mettlen M, Schmid SL. Mino RE, et al. Traffic. 2020 Sep;21(9):603-616. doi: 10.1111/tra.12755. Epub 2020 Jul 22. Traffic. 2020. PMID: 32657003 Free PMC article.
Targeting signals and subunit interactions in coated vesicle adaptor complexes.
Page LJ, Robinson MS. Page LJ, et al. J Cell Biol. 1995 Nov;131(3):619-30. doi: 10.1083/jcb.131.3.619. J Cell Biol. 1995. PMID: 7593184 Free PMC article.

References

1. Science. 1991 Nov 22;254(5035):1197-9 - PubMed
1. J Cell Biol. 1993 Jan;120(1):47-54 - PubMed
1. J Cell Biol. 1989 Mar;108(3):833-42 - PubMed
1. J Biol Chem. 1990 Sep 25;265(27):16514-20 - PubMed
1. Curr Opin Cell Biol. 1991 Aug;3(4):634-41 - PubMed

Assembly and targeting of adaptin chimeras in transfected cells - PubMed (original) (raw)