A genome-wide resource for the analysis of protein localisation in Drosophila

Mihail Sarov; Christiane Barz; Helena Jambor; Marco Y Hein; Christopher Schmied; Dana Suchold; Bettina Stender; Stephan Janosch; Vinay Vikas K J; R T Krishnan; Aishwarya Krishnamoorthy; Irene R S Ferreira; Radoslaw K Ejsmont; Katja Finkl; Susanne Hasse; Philipp Kämpfer; Nicole Plewka; Elisabeth Vinis; Siegfried Schloissnig; Elisabeth Knust; Volker Hartenstein; Matthias Mann; Mani Ramaswami; K VijayRaghavan; Pavel Tomancak; Frank Schnorrer

doi:10.7554/elife.12068

A genome-wide resource for the analysis of protein localisation in Drosophila

Mihail Sarov, Christiane Barz, Helena Jambor, Marco Y Hein, Christopher Schmied, Dana Suchold, Bettina Stender, Stephan Janosch, Vinay Vikas K J, R T Krishnan, Aishwarya Krishnamoorthy, Irene R S Ferreira, Radoslaw K Ejsmont, Katja Finkl, Susanne Hasse, Philipp Kämpfer, Nicole Plewka, Elisabeth Vinis, Siegfried Schloissnig, Elisabeth KnustVolker Hartenstein, Matthias Mann, Mani Ramaswami, K VijayRaghavan, Pavel Tomancak, Frank Schnorrer

134 Citations (Scopus)

Abstract

The Drosophila genome contains >13000 protein-coding genes, the majority of which remain poorly investigated. Important reasons include the lack of antibodies or reporter constructs to visualise these proteins. Here, we present a genome-wide fosmid library of 10000 GFP-tagged clones, comprising tagged genes and most of their regulatory information. For 880 tagged proteins, we created transgenic lines, and for a total of 207 lines, we assessed protein expression and localisation in ovaries, embryos, pupae or adults by stainings and live imaging approaches. Importantly, we visualised many proteins at endogenous expression levels and found a large fraction of them localising to subcellular compartments. By applying genetic complementation tests, we estimate that about two-thirds of the tagged proteins are functional. Moreover, these tagged proteins enable interaction proteomics from developing pupae and adult flies. Taken together, this resource will boost systematic analysis of protein expression and localisation in various cellular and developmental contexts.

Original language	English
Journal	eLife
Volume	5
Pages (from-to)	e12068
ISSN	2050-084X
DOIs	https://doi.org/10.7554/elife.12068
Publication status	Published - 20 Feb 2016
Externally published	Yes

Keywords

Animal Structures
Animals
Animals, Genetically Modified
Drosophila
Drosophila Proteins
Entomology
Gene Library
Genes, Reporter
Genome, Insect
Green Fluorescent Proteins
Image Processing, Computer-Assisted
Molecular Biology
Optical Imaging
Recombinant Fusion Proteins
Staining and Labeling
Journal Article
Research Support, Non-U.S. Gov't

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http://europepmc.org/articles/pmc4805545?pdf=renderLicence: CC BY

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Sarov, M., Barz, C., Jambor, H., Hein, M. Y., Schmied, C., Suchold, D., Stender, B., Janosch, S., K J, V. V., Krishnan, R. T., Krishnamoorthy, A., Ferreira, I. R. S., Ejsmont, R. K., Finkl, K., Hasse, S., Kämpfer, P., Plewka, N., Vinis, E., Schloissnig, S., ... Schnorrer, F. (2016). A genome-wide resource for the analysis of protein localisation in Drosophila. eLife, 5, e12068. https://doi.org/10.7554/elife.12068

Sarov, M, Barz, C, Jambor, H, Hein, MY, Schmied, C, Suchold, D, Stender, B, Janosch, S, K J, VV, Krishnan, RT, Krishnamoorthy, A, Ferreira, IRS, Ejsmont, RK, Finkl, K, Hasse, S, Kämpfer, P, Plewka, N, Vinis, E, Schloissnig, S, Knust, E, Hartenstein, V, Mann, M, Ramaswami, M, VijayRaghavan, K, Tomancak, P & Schnorrer, F 2016, 'A genome-wide resource for the analysis of protein localisation in Drosophila', eLife, vol. 5, pp. e12068. https://doi.org/10.7554/elife.12068

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abstract = "The Drosophila genome contains >13000 protein-coding genes, the majority of which remain poorly investigated. Important reasons include the lack of antibodies or reporter constructs to visualise these proteins. Here, we present a genome-wide fosmid library of 10000 GFP-tagged clones, comprising tagged genes and most of their regulatory information. For 880 tagged proteins, we created transgenic lines, and for a total of 207 lines, we assessed protein expression and localisation in ovaries, embryos, pupae or adults by stainings and live imaging approaches. Importantly, we visualised many proteins at endogenous expression levels and found a large fraction of them localising to subcellular compartments. By applying genetic complementation tests, we estimate that about two-thirds of the tagged proteins are functional. Moreover, these tagged proteins enable interaction proteomics from developing pupae and adult flies. Taken together, this resource will boost systematic analysis of protein expression and localisation in various cellular and developmental contexts.",

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AU - Sarov, Mihail

AU - Barz, Christiane

AU - Jambor, Helena

AU - Hein, Marco Y

AU - Schmied, Christopher

AU - Suchold, Dana

AU - Stender, Bettina

AU - Janosch, Stephan

AU - K J, Vinay Vikas

AU - Krishnan, R T

AU - Krishnamoorthy, Aishwarya

AU - Ferreira, Irene R S

AU - Ejsmont, Radoslaw K

AU - Finkl, Katja

AU - Hasse, Susanne

AU - Kämpfer, Philipp

AU - Plewka, Nicole

AU - Vinis, Elisabeth

AU - Schloissnig, Siegfried

AU - Knust, Elisabeth

AU - Hartenstein, Volker

AU - Mann, Matthias

AU - Ramaswami, Mani

AU - VijayRaghavan, K

AU - Tomancak, Pavel

AU - Schnorrer, Frank

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N2 - The Drosophila genome contains >13000 protein-coding genes, the majority of which remain poorly investigated. Important reasons include the lack of antibodies or reporter constructs to visualise these proteins. Here, we present a genome-wide fosmid library of 10000 GFP-tagged clones, comprising tagged genes and most of their regulatory information. For 880 tagged proteins, we created transgenic lines, and for a total of 207 lines, we assessed protein expression and localisation in ovaries, embryos, pupae or adults by stainings and live imaging approaches. Importantly, we visualised many proteins at endogenous expression levels and found a large fraction of them localising to subcellular compartments. By applying genetic complementation tests, we estimate that about two-thirds of the tagged proteins are functional. Moreover, these tagged proteins enable interaction proteomics from developing pupae and adult flies. Taken together, this resource will boost systematic analysis of protein expression and localisation in various cellular and developmental contexts.

AB - The Drosophila genome contains >13000 protein-coding genes, the majority of which remain poorly investigated. Important reasons include the lack of antibodies or reporter constructs to visualise these proteins. Here, we present a genome-wide fosmid library of 10000 GFP-tagged clones, comprising tagged genes and most of their regulatory information. For 880 tagged proteins, we created transgenic lines, and for a total of 207 lines, we assessed protein expression and localisation in ovaries, embryos, pupae or adults by stainings and live imaging approaches. Importantly, we visualised many proteins at endogenous expression levels and found a large fraction of them localising to subcellular compartments. By applying genetic complementation tests, we estimate that about two-thirds of the tagged proteins are functional. Moreover, these tagged proteins enable interaction proteomics from developing pupae and adult flies. Taken together, this resource will boost systematic analysis of protein expression and localisation in various cellular and developmental contexts.

KW - Animal Structures

KW - Animals

KW - Animals, Genetically Modified

KW - Drosophila

KW - Drosophila Proteins

KW - Entomology

KW - Gene Library

KW - Genes, Reporter

KW - Genome, Insect

KW - Green Fluorescent Proteins

KW - Image Processing, Computer-Assisted

KW - Molecular Biology

KW - Optical Imaging

KW - Recombinant Fusion Proteins

KW - Staining and Labeling

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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DO - 10.7554/elife.12068

M3 - Journal article

C2 - 26896675

SN - 2050-084X

VL - 5

SP - e12068

JO - eLife

JF - eLife

ER -

A genome-wide resource for the analysis of protein localisation in Drosophila

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Keywords

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