Subcellular localisations of the CPTI collection of YFP-tagged proteins inDrosophilaembryos

Abstract

A key challenge in the post-genomic area is to identify the function of the genes discovered, with many still uncharacterised in all metazoans. A first step is transcription pattern characterisation, for which we now have near whole-genome coverage in Drosophila. However, we have much more limited information about the expression and subcellular localisation of the corresponding proteins. The Cambridge Protein Trap Consortium generated, via piggyBac transposition, over 600 novel YFP-trap proteins tagging just under 400 Drosophila loci. Here, we characterise the subcellular localisations and expression patterns of these insertions, called the CPTI lines, in Drosophila embryos. We have systematically analysed subcellular localisations at cellularisation (stage 5) and recorded expression patterns at stage 5, at mid-embryogenesis (stage 11) and at late embryogenesis (stages 15-17). At stage 5, 31% of the nuclear lines (41) and 26% of the cytoplasmic lines (67) show discrete localisations that provide clues on the function of the protein and markers for organelles or regions, including nucleoli, the nuclear envelope, nuclear speckles, centrosomes, mitochondria, the endoplasmic reticulum, Golgi, lysosomes and peroxisomes. We characterised the membranous/cortical lines (102) throughout stage 5 to 10 during epithelial morphogenesis, documenting their apico-basal position and identifying those secreted in the extracellular space. We identified the tricellular vertices as a specialized membrane domain marked by the integral membrane protein Sidekick. Finally, we categorised the localisation of the membranous/cortical proteins during cytokinesis.

Inhibiting the Mitochondrial Calcium Uniporter during Development Impairs Memory in Adult Drosophila

“…The Drosophila gene CG1876 is the homolog of mammalian MCU and its protein product localizes to mitochondria (Lye et al, 2014). CG18769 was first identified as a candidate gene in a pan-neuronal RNAi screen aimed at discovering new genes that are critical for memory formation (Walkinshaw et al, 2015).…”

Fat2 and Lar Define a Basally Localized Planar Signaling System Controlling Collective Cell Migration

“…Lar 13.2 , FRT40 is from Bateman et al, 2001. Traffic jam-Gal4 (104055), UAS-Abi-RNAi (9749R-3), and msn-YFP (115454) (Lye et al, 2014) are from the Drosophila Genetic Resource Center in Kyoto. sqh-GFP (318484) is from the Vienna Drosophila Resource Center (Sarov et al, 2016).…”

Transient opening of tricellular vertices controls paracellular transport through the follicle epithelium duringDrosophilaoogenesis

“…UAS-E-CadΔC::αCat (67415), UAS-Fas2A(PEST+), UAS-Fas2A(PEST-) ( Silies and Klambt, 2010), Gli-YFP (Lye et al, 2014), E-Cad::3xGFP, E-Cad::3xmTagRFP, MyoII::3xGFP (Pinheiro et al, 2017), Vinculin-GFP (Maartens et al, 2016), UAS-Shi-DN (5811), UAS-MLCK (37528; Kim et al, 2002), UAS-Rok.CAT (6669; Winter et al, 2001), hs-Flp 122 , aka L200 FRT40A (Byri et al, 2015), FRTG13 shg 1 , FRTG13 shg 2IH , ubi-nlsGFP FRT40A (5189), FRTG13 ubi-nlsGFP (5826). Oregon R was used as the wild-type strain.…”