NECAPs are negative regulators of the AP2 clathrin adaptor complex
Eukaryotic cells internalize transmembrane receptors via clathrin-mediated endocytosis, but it remains unclear how the machinery underpinning this process is regulated. We recently discovered that membrane-associated muniscin proteins such as FCHo and SGIP initiate endocytosis by converting the AP2...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2018-01-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/32242 |
| _version_ | 1811181435452653568 |
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| author | Gwendolyn M Beacham Edward A Partlow Jeffrey J Lange Gunther Hollopeter |
| author_facet | Gwendolyn M Beacham Edward A Partlow Jeffrey J Lange Gunther Hollopeter |
| author_sort | Gwendolyn M Beacham |
| collection | DOAJ |
| description | Eukaryotic cells internalize transmembrane receptors via clathrin-mediated endocytosis, but it remains unclear how the machinery underpinning this process is regulated. We recently discovered that membrane-associated muniscin proteins such as FCHo and SGIP initiate endocytosis by converting the AP2 clathrin adaptor complex to an open, active conformation that is then phosphorylated (Hollopeter et al., 2014). Here we report that loss of ncap-1, the sole C. elegans gene encoding an adaptiN Ear-binding Coat-Associated Protein (NECAP), bypasses the requirement for FCHO-1. Biochemical analyses reveal AP2 accumulates in an open, phosphorylated state in ncap-1 mutant worms, suggesting NECAPs promote the closed, inactive conformation of AP2. Consistent with this model, NECAPs preferentially bind open and phosphorylated forms of AP2 in vitro and localize with constitutively open AP2 mutants in vivo. NECAPs do not associate with phosphorylation-defective AP2 mutants, implying that phosphorylation precedes NECAP recruitment. We propose NECAPs function late in endocytosis to inactivate AP2. |
| first_indexed | 2024-04-11T09:16:45Z |
| format | Article |
| id | doaj.art-173d1e462426453eb665ef20faeffdbd |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:16:45Z |
| publishDate | 2018-01-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-173d1e462426453eb665ef20faeffdbd2022-12-22T04:32:18ZengeLife Sciences Publications LtdeLife2050-084X2018-01-01710.7554/eLife.32242NECAPs are negative regulators of the AP2 clathrin adaptor complexGwendolyn M Beacham0Edward A Partlow1Jeffrey J Lange2Gunther Hollopeter3https://orcid.org/0000-0002-6409-0530Department of Molecular Medicine, Cornell University, Ithaca, United StatesDepartment of Molecular Medicine, Cornell University, Ithaca, United StatesStowers Institute for Medical Research, Kansas City, United StatesDepartment of Molecular Medicine, Cornell University, Ithaca, United StatesEukaryotic cells internalize transmembrane receptors via clathrin-mediated endocytosis, but it remains unclear how the machinery underpinning this process is regulated. We recently discovered that membrane-associated muniscin proteins such as FCHo and SGIP initiate endocytosis by converting the AP2 clathrin adaptor complex to an open, active conformation that is then phosphorylated (Hollopeter et al., 2014). Here we report that loss of ncap-1, the sole C. elegans gene encoding an adaptiN Ear-binding Coat-Associated Protein (NECAP), bypasses the requirement for FCHO-1. Biochemical analyses reveal AP2 accumulates in an open, phosphorylated state in ncap-1 mutant worms, suggesting NECAPs promote the closed, inactive conformation of AP2. Consistent with this model, NECAPs preferentially bind open and phosphorylated forms of AP2 in vitro and localize with constitutively open AP2 mutants in vivo. NECAPs do not associate with phosphorylation-defective AP2 mutants, implying that phosphorylation precedes NECAP recruitment. We propose NECAPs function late in endocytosis to inactivate AP2.https://elifesciences.org/articles/32242AP2cell biologyclathrin adaptor complexFCHoendocytosisNECAP |
| spellingShingle | Gwendolyn M Beacham Edward A Partlow Jeffrey J Lange Gunther Hollopeter NECAPs are negative regulators of the AP2 clathrin adaptor complex eLife AP2 cell biology clathrin adaptor complex FCHo endocytosis NECAP |
| title | NECAPs are negative regulators of the AP2 clathrin adaptor complex |
| title_full | NECAPs are negative regulators of the AP2 clathrin adaptor complex |
| title_fullStr | NECAPs are negative regulators of the AP2 clathrin adaptor complex |
| title_full_unstemmed | NECAPs are negative regulators of the AP2 clathrin adaptor complex |
| title_short | NECAPs are negative regulators of the AP2 clathrin adaptor complex |
| title_sort | necaps are negative regulators of the ap2 clathrin adaptor complex |
| topic | AP2 cell biology clathrin adaptor complex FCHo endocytosis NECAP |
| url | https://elifesciences.org/articles/32242 |
| work_keys_str_mv | AT gwendolynmbeacham necapsarenegativeregulatorsoftheap2clathrinadaptorcomplex AT edwardapartlow necapsarenegativeregulatorsoftheap2clathrinadaptorcomplex AT jeffreyjlange necapsarenegativeregulatorsoftheap2clathrinadaptorcomplex AT guntherhollopeter necapsarenegativeregulatorsoftheap2clathrinadaptorcomplex |