Function and regulation of Rab3D in the preosteoclast cell line, RAW264.7

April Millar

    Research output: ThesisMaster's Thesis

    77 Downloads (Pure)

    Abstract

    [Truncated] The small GTPases of the Ras family are present at distinct trafficking pathways during vesicle transport. Bone resorption in osteoclasts requires the formation of the ruffled border domain by accelerated fusion of intracellular vesicles to the plasma membrane, an event comparable with vesicle trafficking in other mammalian cells. The Rab3 subfamily consists of four isoforms, Rab3A, Rab3B, Rab3C and Rab3D, which are involved in the mediation of exocytic vesicle transport. This thesis sought to clarify the mechanism of vesicle trafficking mediated by Rab3D in the osteoclast precursor cellline, RAW264.7.

    Using RT-PCR analysis it was demonstrated that RAW264.7 cells expressed both Rab3C and Rab3D. Rab3D was considered the dominant isoform in RAW264.7 cells and was shown to be upregulated during osteoclastogenesis. To further examine the subcellular localisation and role of Rab3D in intracellular trafficking in RAW264.7 cells confocal microscopy and GFP protein techniques were used. Rab3D was shown to localise to pleomorphic vesicular compartments in the cytoplasm of the RAW264.7 cells that associate with microtubules as well as the trans-Golgi network. The expression of three Rab3D mutants, Q81L, N135I and CXC , in RAW264.7 cells showed the importance of prenylation and GTP/GDP cycling in Rab proteins.
    Original languageEnglish
    QualificationMasters
    Awarding Institution
    • The University of Western Australia
    DOIs
    Publication statusUnpublished - 2002

    Take-down notice

    • This thesis has been made available in the UWA Profiles and Research Repository as part of a UWA Library project to digitise and make available theses completed before 2003. If you are the author of this thesis and would like it removed from the UWA Profiles and Research Repository, please contact [email protected]

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