Spatial dynamics of receptor-mediated endocytic trafficking in budding yeast revealed by using fluorescent α-factor derivatives

Junko Y. Toshima, Jiro Toshima, Marko Kaksonen, Adam C. Martin, David S. King, David G. Drubin

    Research output: Contribution to journalArticle

    120 Citations (Scopus)

    Abstract

    Much progress defining the order and timing of endocytic internalization events has come as a result of real-time, live-cell fluorescence microscopy. Although the availability of numerous endocytic mutants makes yeast an especially valuable organism for functional analysis of endocytic dynamics, a serious limitation has been the lack of a fluorescent cargo for receptor-mediated endocytosis. We have now synthesized biologically active fluorescent mating-pheromone derivatives and demonstrated that receptor-mediated endocytosis in budding yeast occurs via the clathrin- and actin-mediated endocytosis pathway. We found that endocytic proteins first assemble into patches on the plasma membrane, and then α-factor associates with the patches. Internalization occurs next, concomitant with actin assembly at patches. Additionally, endocytic vesicles move toward early endosomes on actin cables. Early endosomes also associate with actin cables, and they actively move toward endocytic sites to capture vesicles being released from the plasma membrane. Thus, endocytic vesicle formation and capture of the newly released vesicles by early endosomes occur in a highly concerted manner, mediated by the actin cytoskeleton.

    Original languageEnglish
    Pages (from-to)5793-5798
    Number of pages6
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume103
    Issue number15
    DOIs
    Publication statusPublished - 2006 Apr 11

    Fingerprint

    Saccharomycetales
    Actins
    Endosomes
    Endocytosis
    Transport Vesicles
    Cell Membrane
    Clathrin
    Pheromones
    Actin Cytoskeleton
    Fluorescence Microscopy
    Yeasts
    Proteins

    Keywords

    • Actin
    • Cytoskeleton
    • Endocytosis
    • Endosome

    ASJC Scopus subject areas

    • Genetics
    • General

    Cite this

    Spatial dynamics of receptor-mediated endocytic trafficking in budding yeast revealed by using fluorescent α-factor derivatives. / Toshima, Junko Y.; Toshima, Jiro; Kaksonen, Marko; Martin, Adam C.; King, David S.; Drubin, David G.

    In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, No. 15, 11.04.2006, p. 5793-5798.

    Research output: Contribution to journalArticle

    Toshima, Junko Y. ; Toshima, Jiro ; Kaksonen, Marko ; Martin, Adam C. ; King, David S. ; Drubin, David G. / Spatial dynamics of receptor-mediated endocytic trafficking in budding yeast revealed by using fluorescent α-factor derivatives. In: Proceedings of the National Academy of Sciences of the United States of America. 2006 ; Vol. 103, No. 15. pp. 5793-5798.
    @article{9f2b2cadc2e9458badf95891bc8be324,
    title = "Spatial dynamics of receptor-mediated endocytic trafficking in budding yeast revealed by using fluorescent α-factor derivatives",
    abstract = "Much progress defining the order and timing of endocytic internalization events has come as a result of real-time, live-cell fluorescence microscopy. Although the availability of numerous endocytic mutants makes yeast an especially valuable organism for functional analysis of endocytic dynamics, a serious limitation has been the lack of a fluorescent cargo for receptor-mediated endocytosis. We have now synthesized biologically active fluorescent mating-pheromone derivatives and demonstrated that receptor-mediated endocytosis in budding yeast occurs via the clathrin- and actin-mediated endocytosis pathway. We found that endocytic proteins first assemble into patches on the plasma membrane, and then α-factor associates with the patches. Internalization occurs next, concomitant with actin assembly at patches. Additionally, endocytic vesicles move toward early endosomes on actin cables. Early endosomes also associate with actin cables, and they actively move toward endocytic sites to capture vesicles being released from the plasma membrane. Thus, endocytic vesicle formation and capture of the newly released vesicles by early endosomes occur in a highly concerted manner, mediated by the actin cytoskeleton.",
    keywords = "Actin, Cytoskeleton, Endocytosis, Endosome",
    author = "Toshima, {Junko Y.} and Jiro Toshima and Marko Kaksonen and Martin, {Adam C.} and King, {David S.} and Drubin, {David G.}",
    year = "2006",
    month = "4",
    day = "11",
    doi = "10.1073/pnas.0601042103",
    language = "English",
    volume = "103",
    pages = "5793--5798",
    journal = "Proceedings of the National Academy of Sciences of the United States of America",
    issn = "0027-8424",
    number = "15",

    }

    TY - JOUR

    T1 - Spatial dynamics of receptor-mediated endocytic trafficking in budding yeast revealed by using fluorescent α-factor derivatives

    AU - Toshima, Junko Y.

    AU - Toshima, Jiro

    AU - Kaksonen, Marko

    AU - Martin, Adam C.

    AU - King, David S.

    AU - Drubin, David G.

    PY - 2006/4/11

    Y1 - 2006/4/11

    N2 - Much progress defining the order and timing of endocytic internalization events has come as a result of real-time, live-cell fluorescence microscopy. Although the availability of numerous endocytic mutants makes yeast an especially valuable organism for functional analysis of endocytic dynamics, a serious limitation has been the lack of a fluorescent cargo for receptor-mediated endocytosis. We have now synthesized biologically active fluorescent mating-pheromone derivatives and demonstrated that receptor-mediated endocytosis in budding yeast occurs via the clathrin- and actin-mediated endocytosis pathway. We found that endocytic proteins first assemble into patches on the plasma membrane, and then α-factor associates with the patches. Internalization occurs next, concomitant with actin assembly at patches. Additionally, endocytic vesicles move toward early endosomes on actin cables. Early endosomes also associate with actin cables, and they actively move toward endocytic sites to capture vesicles being released from the plasma membrane. Thus, endocytic vesicle formation and capture of the newly released vesicles by early endosomes occur in a highly concerted manner, mediated by the actin cytoskeleton.

    AB - Much progress defining the order and timing of endocytic internalization events has come as a result of real-time, live-cell fluorescence microscopy. Although the availability of numerous endocytic mutants makes yeast an especially valuable organism for functional analysis of endocytic dynamics, a serious limitation has been the lack of a fluorescent cargo for receptor-mediated endocytosis. We have now synthesized biologically active fluorescent mating-pheromone derivatives and demonstrated that receptor-mediated endocytosis in budding yeast occurs via the clathrin- and actin-mediated endocytosis pathway. We found that endocytic proteins first assemble into patches on the plasma membrane, and then α-factor associates with the patches. Internalization occurs next, concomitant with actin assembly at patches. Additionally, endocytic vesicles move toward early endosomes on actin cables. Early endosomes also associate with actin cables, and they actively move toward endocytic sites to capture vesicles being released from the plasma membrane. Thus, endocytic vesicle formation and capture of the newly released vesicles by early endosomes occur in a highly concerted manner, mediated by the actin cytoskeleton.

    KW - Actin

    KW - Cytoskeleton

    KW - Endocytosis

    KW - Endosome

    UR - http://www.scopus.com/inward/record.url?scp=33645801592&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=33645801592&partnerID=8YFLogxK

    U2 - 10.1073/pnas.0601042103

    DO - 10.1073/pnas.0601042103

    M3 - Article

    C2 - 16574772

    AN - SCOPUS:33645801592

    VL - 103

    SP - 5793

    EP - 5798

    JO - Proceedings of the National Academy of Sciences of the United States of America

    JF - Proceedings of the National Academy of Sciences of the United States of America

    SN - 0027-8424

    IS - 15

    ER -