A fundamental feature of eukaryotic cells is the secretory pathway that makes and delivers all secreted and membrane proteins. These proteins are inserted and assembled in the endoplasmic reticulum, and then moved through to Golgi apparatus from where they are sorted for delivery to the cell surface or internal organelles. Such membrane traffic is of particular importance for the function of neurons with numerous neurotransmitters and receptors having to be delivered to the correct location in these large and complex cells.
In the cell body of neurons there is central Golgi apparatus which sorts numerous proteins destined for the axons or the dendrites. However, in addition, there are “Golgi outposts” along the length of the dendrites. These outposts are important for not only the delivery of key membrane proteins but also the organisation of microtubules and hence neuronal morphology. However, it is unclear how these outposts differ in composition, structure and function compared to the Golgis found in the cell body of the neuron or in other cell types.
The PhD project will make use of the tractable genetics of Drosophila melanogaster to investigate the properties of Golgi outposts and compare them to Golgis in the neuronal cell body and other cell types. We have developed tools and reagents to examine the Golgi in Drosophila, and the project will involve examining the localisation of Golgi proteins and the effects of mutants in these proteins. In addition, it will make use of methods we have developed to examine the content of intra-Golgi transport vesicles by using vesicle tethers attached to mitochondria to capture and isolate these vesicles.
The project will provide the opportunity to learn Drosophila genetics and apply a range of cell biological methods including CRISPR-Cas9 genome engineering and high-resolution light microscopy.
References
Park SY, Muschalik N, Chadwick J and Munro S (2022).
In vivo characterization of Drosophila golgins reveals redundancy and plasticity of vesicle capture at the Golgi apparatus.
Current Biology (online)
Kemal S, Richardson HS, Dyne ED and Fu MM (2022).
ER and Golgi trafficking in axons, dendrites, and glial processes.
Curr Opin Cell Biol 78, 102119.
Riedel F, Galindo A, Muschalik N and Munro S (2018).
The two TRAPP complexes of metazoans have distinct roles and act on different Rab GTPases.
J Cell Biology 217, 601-617.
Riedel F, Gillingham AK, Rosa-Ferreira C, Galindo A and Munro S (2016).
An antibody toolkit for the study of membrane traffic in Drosophila melanogaster.
Biology Open 5, 987-992.
Wong M. and Munro S. (2014)
Membrane trafficking. The specificity of vesicle traffic to the Golgi is encoded in the golgin coiled-coil proteins.
Science 346, 1256898.