GenomeRNAi - a database for RNAi phenotypes and reagents

Phenotype information for gene 318666 (CG31294)

Screen TitleGene IDGene SymbolReagent IDScorePhenotypeFollow Up
Lipid storage
FBgn0051294
-0.63 none no

Reference

COPI complex is a regulator of lipid homeostasis. Beller et al., 2008

Lipid droplets are ubiquitous triglyceride and sterol ester storage organelles required for energy storage homeostasis and biosynthesis. Although little is known about lipid droplet formation and regulation, it is clear that members of the PAT (perilipin, adipocyte differentiation related protein, tail interacting protein of 47 kDa) protein family coat the droplet surface and mediate interactions with lipases that remobilize the stored lipids. We identified key Drosophila candidate genes for lipid droplet regulation by RNA interference (RNAi) screening with an image segmentation-based optical read-out system, and show that these regulatory functions are conserved in the mouse. Those include the vesicle-mediated Coat Protein Complex I (COPI) transport complex, which is required for limiting lipid storage. We found that COPI components regulate the PAT protein composition at the lipid droplet surface, and promote the association of adipocyte triglyceride lipase (ATGL) with the lipid droplet surface to mediate lipolysis. Two compounds known to inhibit COPI function, Exo1 and Brefeldin A, phenocopy COPI knockdowns. Furthermore, RNAi inhibition of ATGL and simultaneous drug treatment indicate that COPI and ATGL function in the same pathway. These data indicate that the COPI complex is an evolutionarily conserved regulator of lipid homeostasis, and highlight an interaction between vesicle transport systems and lipid droplets.

Screen details

Stable Id: GR00002-A-0
Screen title: Lipid storage
Assay: Lipid droplet staining
Method: High content (microscopy)
Scope:
Screen type: Cell-based
Species: Drosophila melanogaster
Biosource: Cell line
Biomodel: Kc167
Library: , DRSC
Reagent type: dsRNA
Score type: B-score
Cutoff: 2.0 / -1.7
Notes:

Cell size and cell-cycle regulation (1)
FBgn0051294
CG31294
AT21457
sp none no

Reference

Identification of pathways regulating cell size and cell-cycle progression by RNAi. BjÓ§rklund et al., 2006

Many high-throughput loss-of-function analyses of the eukaryotic cell cycle have relied on the unicellular yeast species Saccharomyces cerevisiae and Schizosaccharomyces pombe. In multicellular organisms, however, additional control mechanisms regulate the cell cycle to specify the size of the organism and its constituent organs. To identify such genes, here we analysed the effect of the loss of function of 70% of Drosophila genes (including 90% of genes conserved in human) on cell-cycle progression of S2 cells using flow cytometry. To address redundancy, we also targeted genes involved in protein phosphorylation simultaneously with their homologues. We identify genes that control cell size, cytokinesis, cell death and/or apoptosis, and the G1 and G2/M phases of the cell cycle. Classification of the genes into pathways by unsupervised hierarchical clustering on the basis of these phenotypes shows that, in addition to classical regulatory mechanisms such as Myc/Max, Cyclin/Cdk and E2F, cell-cycle progression in S2 cells is controlled by vesicular and nuclear transport proteins, COP9 signalosome activity and four extracellular-signal-regulated pathways (Wnt, p38betaMAPK, FRAP/TOR and JAK/STAT). In addition, by simultaneously analysing several phenotypes, we identify a translational regulator, eIF-3p66, that specifically affects the Cyclin/Cdk pathway activity.

Screen details

Stable Id: GR00048-A-1
Screen title: Cell size and cell-cycle regulation (1)
Assay: Cell size, DNA content and viability
Method: Flow cytometry
Scope: Kinases, phosphatases and selected genes
Screen type: Cell-based
Species: Drosophila melanogaster
Biosource: Cell line
Biomodel: S2
Library: Custom-made, DGC1, DGC2 and PHOSPHO
Reagent type: dsRNA
Score type: Complex, sp
Cutoff: Complex criteria
Notes: Additional information about the primary sccreen (pooled library) and a secondary screen (number of binucleate cells)

Lipid storage
FBgn0051294
-1.11 none no

Reference

COPI complex is a regulator of lipid homeostasis. Beller et al., 2008

Lipid droplets are ubiquitous triglyceride and sterol ester storage organelles required for energy storage homeostasis and biosynthesis. Although little is known about lipid droplet formation and regulation, it is clear that members of the PAT (perilipin, adipocyte differentiation related protein, tail interacting protein of 47 kDa) protein family coat the droplet surface and mediate interactions with lipases that remobilize the stored lipids. We identified key Drosophila candidate genes for lipid droplet regulation by RNA interference (RNAi) screening with an image segmentation-based optical read-out system, and show that these regulatory functions are conserved in the mouse. Those include the vesicle-mediated Coat Protein Complex I (COPI) transport complex, which is required for limiting lipid storage. We found that COPI components regulate the PAT protein composition at the lipid droplet surface, and promote the association of adipocyte triglyceride lipase (ATGL) with the lipid droplet surface to mediate lipolysis. Two compounds known to inhibit COPI function, Exo1 and Brefeldin A, phenocopy COPI knockdowns. Furthermore, RNAi inhibition of ATGL and simultaneous drug treatment indicate that COPI and ATGL function in the same pathway. These data indicate that the COPI complex is an evolutionarily conserved regulator of lipid homeostasis, and highlight an interaction between vesicle transport systems and lipid droplets.

Screen details

Stable Id: GR00002-A-0
Screen title: Lipid storage
Assay: Lipid droplet staining
Method: High content (microscopy)
Scope:
Screen type: Cell-based
Species: Drosophila melanogaster
Biosource: Cell line
Biomodel: Kc167
Library: , DRSC
Reagent type: dsRNA
Score type: B-score
Cutoff: 2.0 / -1.7
Notes:

Reagent information for gene 318666 (CG31294)

Reagent IDTypeLibrary
BKN30725 dsRNA
BKN|Boutros Lab|1|FlyBase release 4, 5|19708|dsRNA|http://b110-wiki.dkfz.de/signaling/wiki/display/rnaiwiki/Drosophila+RNAi+libraries
72330 dsRNA
Open Biosystems|Open Biosystems|1 and 2|FlyBase release 2 - 4|15466|dsRNA|http://www.openbiosystems.com/
DRSC22217 dsRNA
DRSC|Drosophila RNAi Screening Center (DRSC)|2|FlyBase release 2 - 5|42076|dsRNA|http://www.flyrnai.org/
AMB29518 dsRNA
Ambion|Ambion|1|FlyBase release 2|13071|dsRNA|http://www.invitrogen.com/site/us/en/home/brands/ambion.html?CID=fl-ambion
DRSC26865 dsRNA
DRSC|Drosophila RNAi Screening Center (DRSC)|2|FlyBase release 2 - 5|42076|dsRNA|http://www.flyrnai.org/
105842 UAS-IR construct
KK|Vienna Drosophila RNAi Center (VDRC)|null|FlyBase|10714|UAS-IR construct|http://stockcenter.vdrc.at/control/main

Gene information for gene 318666 (CG31294)

Gene:
Alternate gene names:NEST:bs25d10, BcDNA:AT21457, NEST:bs15e12, DmelCG31294
Description:
Chromosome:3R
Locus:
Biotype:protein-coding
Status:live
Entrez Gene ID:
Homologs:0
FLYBASE:
Uniprot:
RefSeq ID: -

GBrowse information for gene 318666 (CG31294)