ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis
Clathrin-mediated endocytosis (CME) is a major route of entry into eukaryotic cells. A core of evolutionarily ancient genes encodes many components of this system but much of our mechanistic understanding of CME is derived from a phylogenetically narrow sampling of a few model organisms. In the para...
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The Company of Biologists
2015
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| Online Access: | https://eprints.nottingham.ac.uk/35362/ |
| _version_ | 1848795060157546496 |
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| author | Manna, Paul T. Gadelha, Catarina Puttick, Amy E. Field, Mark C. |
| author_facet | Manna, Paul T. Gadelha, Catarina Puttick, Amy E. Field, Mark C. |
| author_sort | Manna, Paul T. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Clathrin-mediated endocytosis (CME) is a major route of entry into eukaryotic cells. A core of evolutionarily ancient genes encodes many components of this system but much of our mechanistic understanding of CME is derived from a phylogenetically narrow sampling of a few model organisms. In the parasite Trypanosoma brucei, which is distantly related to the better characterised animals and fungi, exceptionally fast endocytic turnover aids its evasion of the host immune system. Although clathrin is absolutely essential for this process, the adaptor protein complex 2 (AP2) has been secondarily lost, suggesting mechanistic divergence. Here, we characterise two phosphoinositide-binding monomeric clathrin adaptors, T. brucei (Tb)EpsinR and TbCALM, which in trypanosomes are represented by single genes, unlike the expansions present in animals and fungi. Depletion of these gene products reveals essential, but partially redundant, activities in CME. Ultrastructural analysis of TbCALM and TbEpsinR double-knockdown cells demonstrated severe defects to clathrin-coated pit formation and morphology associated with a dramatic inhibition of endocytosis. Depletion of TbCALM alone, however, produced a distinct lysosomal segregation phenotype, indicating an additional non-redundant role for this protein. Therefore, TbEpsinR and TbCALM represent ancient phosphoinositide-binding proteins with distinct and vital roles in AP2-independent endocytosis. |
| first_indexed | 2025-11-14T19:26:05Z |
| format | Article |
| id | nottingham-35362 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:26:05Z |
| publishDate | 2015 |
| publisher | The Company of Biologists |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-353622020-05-04T17:07:26Z https://eprints.nottingham.ac.uk/35362/ ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis Manna, Paul T. Gadelha, Catarina Puttick, Amy E. Field, Mark C. Clathrin-mediated endocytosis (CME) is a major route of entry into eukaryotic cells. A core of evolutionarily ancient genes encodes many components of this system but much of our mechanistic understanding of CME is derived from a phylogenetically narrow sampling of a few model organisms. In the parasite Trypanosoma brucei, which is distantly related to the better characterised animals and fungi, exceptionally fast endocytic turnover aids its evasion of the host immune system. Although clathrin is absolutely essential for this process, the adaptor protein complex 2 (AP2) has been secondarily lost, suggesting mechanistic divergence. Here, we characterise two phosphoinositide-binding monomeric clathrin adaptors, T. brucei (Tb)EpsinR and TbCALM, which in trypanosomes are represented by single genes, unlike the expansions present in animals and fungi. Depletion of these gene products reveals essential, but partially redundant, activities in CME. Ultrastructural analysis of TbCALM and TbEpsinR double-knockdown cells demonstrated severe defects to clathrin-coated pit formation and morphology associated with a dramatic inhibition of endocytosis. Depletion of TbCALM alone, however, produced a distinct lysosomal segregation phenotype, indicating an additional non-redundant role for this protein. Therefore, TbEpsinR and TbCALM represent ancient phosphoinositide-binding proteins with distinct and vital roles in AP2-independent endocytosis. The Company of Biologists 2015-06-01 Article PeerReviewed Manna, Paul T., Gadelha, Catarina, Puttick, Amy E. and Field, Mark C. (2015) ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis. Journal of Cell Science, 128 (11). pp. 2130-2142. ISSN 1477-9137 http://jcs.biologists.org/content/128/11/2130 doi: 10.1242/jcs.167726 doi: 10.1242/jcs.167726 |
| spellingShingle | Manna, Paul T. Gadelha, Catarina Puttick, Amy E. Field, Mark C. ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis |
| title | ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis |
| title_full | ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis |
| title_fullStr | ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis |
| title_full_unstemmed | ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis |
| title_short | ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis |
| title_sort | enth and anth domain proteins participate in ap2-independent clathrin-mediated endocytosis |
| url | https://eprints.nottingham.ac.uk/35362/ https://eprints.nottingham.ac.uk/35362/ https://eprints.nottingham.ac.uk/35362/ |