ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis

Paul T. Manna, Catarina Gadelha, Amy E. Puttick, Mark C. Field (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    16 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)2130-2142
    Number of pages13
    JournalJournal of Cell Science
    Volume128
    Issue number11
    DOIs
    Publication statusPublished - 2015

    Fingerprint

    Adaptor Protein Complex 2
    Clathrin
    Endocytosis
    Trypanosoma brucei brucei
    Phosphatidylinositols
    Fungi
    Vesicular Transport Adaptor Proteins
    Genes
    Trypanosomiasis
    Eukaryotic Cells
    Immune System
    Carrier Proteins
    Parasites
    Protein Domains
    Phenotype

    Keywords

    • Endocytosis
    • Evolution
    • Phosphoinositide
    • Trafficking
    • Trypanosome

    Cite this

    Manna, Paul T. ; Gadelha, Catarina ; Puttick, Amy E. ; Field, Mark C. / ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis. In: Journal of Cell Science. 2015 ; Vol. 128, No. 11. pp. 2130-2142.
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    ENTH and ANTH domain proteins participate in AP2-independent clathrin-mediated endocytosis. / Manna, Paul T.; Gadelha, Catarina; Puttick, Amy E.; Field, Mark C. (Lead / Corresponding author).

    In: Journal of Cell Science, Vol. 128, No. 11, 2015, p. 2130-2142.

    Research output: Contribution to journalArticle

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