The plant Golgi apparatus modifies and sorts incoming proteins from the endoplasmic reticulum (ER), and synthesises cell wall matrix material. Plant cells possess numerous motile Golgi bodies, which are connected to the ER by yet to be identified tethering factors. Previous studies indicated a role of cis-Golgi plant golgins (long coiled-coil domains proteins anchored to Golgi membranes) in Golgi biogenesis. Here we show a tethering role for the golgin AtCASP at the ER-Golgi interface. Using live-cell imaging, Golgi body dynamics were compared in Arabidopsis thaliana leaf epidermal cells expressing fluorescently tagged AtCASP, a truncated AtCASP-ΔCC lacking the coiled-coil domains, and the Golgi marker STtmd. Golgi body speed and displacement were significantly reduced in AtCASP-ΔCC lines. Using a dual-colour optical trapping system and a TIRF-tweezer system, individual Golgi bodies were captured in planta. Golgi bodies in AtCASP-ΔCC lines were easier to trap, and the ER-Golgi connection was more easily disrupted. Occasionally, the ER tubule followed a trapped Golgi body with a gap, indicating the presence of other tethering factors. Our work confirms that the intimate ER-Golgi association can be disrupted or weakened by expression of truncated AtCASP-ΔCC, and suggests that this connection is most likely maintained by a golgin-mediated tethering complex.
Osterrieder, Anne Hawes, Chris Sparkes, IBotchway, SWard, AKetelaar, Tde Ruijter, N
Faculty of Health and Life Sciences\Department of Biological and Medical Sciences
Year of publication: 2017Date of RADAR deposit: 2017-04-27