Analysis of the expression patterns, subcellular localisations and interaction partners of Drosophila proteins using a pigP protein trap library

Abstract

Although we now have a wealth of information on the transcription patterns of all the genes in the Drosophila genome, much less is known about the properties of the encoded proteins. To provide information on the expression patterns and subcellular localisations of many proteins in parallel, we have performed a large-scale protein trap screen using a hybrid piggyBac vector carrying an artificial exon encoding yellow fluorescent protein (YFP) and protein affinity tags. From screening 41 million embryos, we recovered 616 verified independent YFP-positive lines representing protein traps in 374 genes, two-thirds of which had not been tagged in previous P element protein trap screens. Over 20 different research groups then characterized the expression patterns of the tagged proteins in a variety of tissues and at several developmental stages. In parallel, we purified many of the tagged proteins from embryos using the affinity tags and identified co-purifying proteins by mass spectrometry. The fly stocks are publicly available through the Kyoto Drosophila Genetics Resource Center. All our data are available via an open access database (Flannotator), which provides comprehensive information on the expression patterns, subcellular localisations and in vivo interaction partners of the trapped proteins. Our resource substantially increases the number of available protein traps in Drosophila and identifies new markers for cellular organelles and structures.

Tissue tension and not interphase cell shape determines cell division orientation in the Drosophila follicular epithelium

“…The EMBO Journal 38: e100072 | 2019 (Rebollo et al, 2004), Ubi-Abnormal spindles-GFP (Rujano et al, 2013), Ubi-Centrosomin-RFP (Basto et al, 2008), Ubi-Centrosomin-GFP (Conduit et al, 2010), Ubi-Pins-YFP (Bellaïche et al, 2001), Basigin::YFP (Lowe et al, 2014;Bergstralh et al, 2015), GFP-Mud (Bosveld et al, 2016), Pavarotti-KLP::GFP (Minestrini et al, 2003), The following background stocks were used to generate mitotic clones, which were induced by heat shock at 37°for multiple periods of 2 h: RFP-nls, hsflp, FRT19A, and hsflp; FRT40A RFP-nls, and hsflp;; FRT82B RFP-nls. Ectopic expression in follicle cells was driven by Traffic Jam-Gal4 (Olivieri et al, 2010), or actin5c-FLPout-Gal4, UAS-GFP (Fig S4G only).…”

In Vivo Interaction Proteomics in Caenorhabditis elegans Embryos Provides New Insights into P Granule Dynamics

“…On the other hand, stringent purification procedures, such as tandem affinity purification (TAP), reduce nonspecific binding but could lead to loss of transient interactions. Most published studies did not employ quantification, which makes it difficult to distinguish genuine interaction partners from nonspecific contaminants (15)(16)(17)(18). Quantitative affinity purification and mass spectrometry solves this problem by using quantification as an additional filter (19 -21).…”

Mechanical Control of Whole Body Shape by a Single Cuticular Protein Obstructor-E in Drosophila melanogaster

“…Canton-S and w 1118 were used as wild-type. Cambridge Protein Trap Insertions (CPTI) 002377 and 100067 [29] were from the Kyoto Stock Center, UAS- obst-E dsRNA (11142R-1) was from the National Institute of Genetics, and hsFLP , UAS- Dicer2 [30], Actin5C -GAL4 ( Act -GAL4) and Ay GAL4, UAS-GFP.S65T [31] were from the Bloomington Stock Center. FRT-bearing PiggyBac insertions f04201 and f06711 from the Exelixis Collection (Harvard) were used to generate the deletion of four genes including obst-E ( obst-E Del ) by FLP-FRT-based method [32].…”