RGD Reference Report - Microtubule-dependent movement of late endocytic vesicles in vitro: requirements for Dynein and Kinesin. - Rat Genome Database

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Microtubule-dependent movement of late endocytic vesicles in vitro: requirements for Dynein and Kinesin.

Authors: Bananis, E  Nath, S  Gordon, K  Satir, P  Stockert, RJ  Murray, JW  Wolkoff, AW 
Citation: Bananis E, etal., Mol Biol Cell. 2004 Aug;15(8):3688-97. Epub 2004 Jun 4.
RGD ID: 1581596
Pubmed: PMID:15181154   (View Abstract at PubMed)
PMCID: PMC491828   (View Article at PubMed Central)
DOI: DOI:10.1091/mbc.E04-04-0278   (Journal Full-text)

Our previous studies demonstrated that fluorescent early endocytic vesicles prepared from rat liver after injection of Texas red asialoorosomucoid contain asialoglycoprotein and its receptor and move and undergo fission along microtubules using kinesin I and KIFC2, with Rab4 regulating KIFC2 activity (J. Cell Sci. 116, 2749, 2003). In the current study, procedures to prepare fluorescent late endocytic vesicles were devised. In addition, flow cytometry was utilized to prepare highly purified fluorescent endocytic vesicles, permitting validation of microscopy-based experiments as well as direct biochemical analysis. These studies revealed that late vesicles bound to and moved along microtubules, but in contrast to early vesicles, did not undergo fission. As compared with early vesicles, late vesicles had reduced association with receptor, Rab4, and kinesin I but were highly associated with dynein, Rab7, dynactin, and KIF3A. Dynein and KIF3A antibodies inhibited late vesicle motility, whereas kinesin I and KIFC2 antibodies had no effect. Dynamitin antibodies prevented the association of late vesicles with microtubules. These results indicate that acquisition and exchange of specific motor and regulatory proteins characterizes and may regulate the transition of early to late endocytic vesicles. Flow cytometric purification should ultimately facilitate detailed proteomic analysis and mapping of endocytic vesicle-associated proteins.

Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
positive regulation of receptor-mediated endocytosis  IDA 1581596 RGD 

Objects Annotated

Genes (Rattus norvegicus)
Kif3a  (kinesin family member 3a)


Additional Information