DOI: 10.1083/jcb.200107140. Assembly and function of AP-3 complexes in cells expressing mutant subunits

Assembly and function of AP-3 complexes in cells expressing mutant subunits

10.1083/jcb.200107140

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

The mouse mutants mocha and pearl are deficient in the AP-3 δ and β3A subunits, respectively. We have used cells from these mice to investigate both the assembly of AP-3 complexes and AP-3 function. In mocha cells, the β3 and μ3 subunits coassemble into a heterodimer, whereas the σ3 subunit remains monomeric. In pearl cells, the δ and σ3 subunits coassemble into a heterodimer, whereas μ3 gets destroyed. The yeast two hybrid system was used to confirm these interactions, and also to demonstrate that the A (ubiquitous) and B (neuronal-specific) isoforms of β3 and μ3 can interact with each other. Pearl cell lines were generated that express β3A, β3B, a β3Aβ2 chimera, two β3A deletion mutants, and a β3A point mutant lacking a functional clathrin binding site. All six constructs assembled into complexes and were recruited onto membranes. However, only β3A, β3B, and the point mutant gave full functional rescue, as assayed by LAMP-1 sorting. The β3Aβ2 chimera and the β3A short deletion mutant gave partial functional rescue, whereas the β3A truncation mutant gave no functional rescue. These results indicate that the hinge and/or ear domains of β3 are important for function, but the clathrin binding site is not needed.

Bibliography

Peden, A. A., Rudge, R. E., Lui, W. W. Y., & Robinson, M. S. (2002). Assembly and function of AP-3 complexes in cells expressing mutant subunits. The Journal of Cell Biology, 156(2), 327â336.

Authors 4

Andrew A. Peden (first)
Rachel E. Rudge (additional)
Winnie W.Y. Lui (additional)
Margaret S. Robinson (additional)

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 12:46 p.m.)
Deposited	2 years, 1 month ago (July 22, 2023, 9:20 a.m.)
Indexed	1 month, 3 weeks ago (July 2, 2025, 9:30 a.m.)
Issued	23 years, 7 months ago (Jan. 21, 2002)
Published	23 years, 7 months ago (Jan. 21, 2002)
Published Online	23 years, 7 months ago (Jan. 21, 2002)
Published Print	23 years, 7 months ago (Jan. 21, 2002)

Funders 0

None

BibTeX

@article{Peden_2002, title={Assembly and function of AP-3 complexes in cells expressing mutant subunits}, volume={156}, ISSN={0021-9525}, url={http://dx.doi.org/10.1083/jcb.200107140}, DOI={10.1083/jcb.200107140}, number={2}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Peden, Andrew A. and Rudge, Rachel E. and Lui, Winnie W.Y. and Robinson, Margaret S.}, year={2002}, month=jan, pages={327–336} }

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  "abstract": "<jats:p>The mouse mutants mocha and pearl are deficient in the AP-3 \u03b4 and \u03b23A subunits, respectively. We have used cells from these mice to investigate both the assembly of AP-3 complexes and AP-3 function. In mocha cells, the \u03b23 and \u03bc3 subunits coassemble into a heterodimer, whereas the \u03c33 subunit remains monomeric. In pearl cells, the \u03b4 and \u03c33 subunits coassemble into a heterodimer, whereas \u03bc3 gets destroyed. The yeast two hybrid system was used to confirm these interactions, and also to demonstrate that the A (ubiquitous) and B (neuronal-specific) isoforms of \u03b23 and \u03bc3 can interact with each other. Pearl cell lines were generated that express \u03b23A, \u03b23B, a \u03b23A\u03b22 chimera, two \u03b23A deletion mutants, and a \u03b23A point mutant lacking a functional clathrin binding site. All six constructs assembled into complexes and were recruited onto membranes. However, only \u03b23A, \u03b23B, and the point mutant gave full functional rescue, as assayed by LAMP-1 sorting. The \u03b23A\u03b22 chimera and the \u03b23A short deletion mutant gave partial functional rescue, whereas the \u03b23A truncation mutant gave no functional rescue. These results indicate that the hinge and/or ear domains of \u03b23 are important for function, but the clathrin binding site is not needed.</jats:p>",
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