title: 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.
authors.affiliation: The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Nick
authors.initials: N
authors.lastname: Lowe
authors.affiliation: The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK The Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
authors.forename: Johanna S
authors.initials: JS
authors.lastname: Rees
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: John
authors.initials: J
authors.lastname: Roote
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Ed
authors.initials: E
authors.lastname: Ryder
authors.affiliation: The Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
authors.forename: Irina M
authors.initials: IM
authors.lastname: Armean
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Glynnis
authors.initials: G
authors.lastname: Johnson
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Emma
authors.initials: E
authors.lastname: Drummond
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Helen
authors.initials: H
authors.lastname: Spriggs
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Jenny
authors.initials: J
authors.lastname: Drummond
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Jose P
authors.initials: JP
authors.lastname: Magbanua
authors.affiliation: The Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Huw
authors.initials: H
authors.lastname: Naylor
authors.affiliation: The Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Sanson
authors.affiliation: The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Bastock
authors.affiliation: The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Sven
authors.initials: S
authors.lastname: Huelsmann
authors.affiliation: The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Vitor
authors.initials: V
authors.lastname: Trovisco
authors.affiliation: The Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK.
authors.forename: Matthias
authors.initials: M
authors.lastname: Landgraf
authors.affiliation: Institute for Adaptive and Neural Computation, University of Edinburgh, 10 Crichton Street, Edinburgh EH8 9AB, UK.
authors.forename: Seymour
authors.initials: S
authors.lastname: Knowles-Barley
authors.affiliation: Institute for Adaptive and Neural Computation, University of Edinburgh, 10 Crichton Street, Edinburgh EH8 9AB, UK.
authors.forename: J Douglas
authors.initials: JD
authors.lastname: Armstrong
authors.affiliation: Cardiff School of Biosciences, The Sir Martin Evans Building, Museum Avenue, Cardiff CF10 3AX, UK.
authors.forename: Helen
authors.initials: H
authors.lastname: White-Cooper
authors.affiliation: Department of Genetics, University of Leicester, Adrian Building, University Road, Leicester LE1 7RH, UK.
authors.forename: Celia
authors.initials: C
authors.lastname: Hansen
authors.affiliation: Centre for Advanced Microscopy, University of Sussex, School of Life Sciences, John Maynard Smith Building, Falmer, Brighton and Hove BN1 9QG, UK.
authors.forename: Roger G
authors.initials: RG
authors.lastname: Phillips
authors.affiliation: The Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
authors.forename: Kathryn S
authors.initials: KS
authors.lastname: Lilley
authors.affiliation: The Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.
authors.forename: Steven
authors.initials: S
authors.lastname: Russell
authors.affiliation: The Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK d.stjohnston@gurdon.cam.ac.uk.
authors.forename: Daniel
authors.initials: D
authors.lastname: St Johnston
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry; D003433:Crosses, Genetic; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000502:physiology*; D005091:Exons; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005821:Genetic Techniques; D016678:Genome; D008164:Luminescent Proteins / Q000737:chemistry; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D010053:Ovary / Q000378:metabolism; D012737:Sex Factors; D013737:Testis / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D029721:Drosophila Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; C054562:yellow fluorescent protein, Bacteria
keywords: Affinity purification; Cytoophidia; Live imaging; Protein trap; piggyBac