Biochemistry and Pharmacology - Research Publications

Permanent URI for this collection

http://hdl.handle.net/11343/214

Filters

2019 - 2019 1 2020 - 2022 1
2
Reset filters

Settings
Statistics
Citations
Author: Gleeson, Paul × Author: Makhoul, Christian × Author: Williamson, Nicholas × Start date: 2010 ×

Search Results

Now showing 1 - 2 of 2
  • Item
    Thumbnail Image
    Interacting partners of Golgi-localized small G protein Arl5b identified by a combination of in vivo proximity labelling and GFP-Trap pull down
    Houghton, FJ ; Makhoul, C ; Cho, EH-J ; Williamson, NA ; Gleeson, PA (WILEY, 2022-09)
    The small G protein Arl5b is localised on the trans-Golgi network (TGN) and regulates endosomes-to-TGN transport. Here, we combined in vivo and in vitro techniques to map the interactive partners and near neighbours of Arl5b at the TGN, using constitutively active, membrane-bound Arl5b(Q70L)-GFP in stably expressing HeLa cells, and the proximity labelling techniques BioID and APEX2 in parallel with GFP-Trap pull down. From MS analysis, 22 Golgi proteins were identified; 50% were TGN-localised Rabs, Arfs and Arls. The scaffold/tethering factors ACBD3 (GCP60) and PIST (GOPC) were also identified, and we show that Arl5b is required for TGN recruitment of ACBD3. Overall, the combination of in vivo labelling and direct pull downs indicates a highly organised complex of small G proteins on TGN membranes.
  • Item
    Thumbnail Image
    Intersectin-1 interacts with the golgin GCC88 to couple the actin network and Golgi architecture
    Makhoul, C ; Gosavi, P ; Duffield, R ; Delbridge, B ; Williamson, NA ; Gleeson, PA ; Glick, BS (AMER SOC CELL BIOLOGY, 2019-02-01)
    The maintenance of the Golgi ribbon relies on a dynamic balance between the actin and microtubule networks; however, the pathways controlling actin networks remain poorly defined. Previously, we showed that the trans-Golgi network (TGN) membrane tether/golgin, GCC88, modulates the Golgi ribbon architecture. Here, we show that dispersal of the Golgi ribbon by GCC88 is dependent on actin and the involvement of nonmuscle myosin IIA. We have identified the long isoform of intersectin-1 (ITSN-1), a guanine nucleotide exchange factor for Cdc42, as a novel Golgi component and an interaction partner of GCC88 responsible for mediating the actin-dependent dispersal of the Golgi ribbon. We show that perturbation of Golgi morphology by changes in membrane flux, mediated by silencing the retromer subunit Vps26, or in a model of neurodegeneration, induced by Tau overexpression, are also dependent on the ITSN-1-GCC88 interaction. Overall, our study reveals a role for a TGN golgin and ITSN-1 in linking to the actin cytoskeleton and regulating the balance between a compact Golgi ribbon and a dispersed Golgi, a pathway with relevance to pathophysiological conditions.