Regulation of growth and TORC1 signalling by the novel evolutionarily conserved amino acid transporter, SLC38A10: a new player in amino acid sensing?. (360G-Wellcome-093326_Z_10_Z)

We will dissect out the functions and mechanism of action of the growth regulatory SLC38A10 transporter in Drosophila, using the combination of genetic and molecular genetic approaches we have developed in our pioneering studies of amino acid sensing by the PATs. We will employ analysis of mutant animals and somatic mutant clones, RNAi and overexpression studies to determine in which tissues SLC38A10 controls growth (we already know it affects growth in the eye and a viable mutant appears to mod ulate growth in all parts of the adult). This transporter is expressed ubiquitously, but we will generate anti-SLC38A10 antisera to assess this its subcellular localisation and shuttling, properties that are fundamental to the regulation of the PAT amino acid sensing mechanism. We will determine the transport and TORC1-regulatory properties of SLC38A10, using a combination of studies in Drosophila S2 cell culture and microinjected Xenopus oocytes. This will then allow us to design cell culture a nd in vivo overexpression/rescue experiments to test whether SLC38A10 activates TORC1 through amino acid transport or through a so-called transceptor (signalling) mechanism, and to elucidate which features that distinguish SLC38A10 transporters from all other AATs are critical to this molecule s functions.