Project description:Transcriptome and Translatome in Drosophila S2 cells treated by tsRNA transfection, serum deprivation, ago 2 knockdown and rapamycin
Project description:Transfer RNA-derived small RNAs (tsRNAs) are an emerging class of small RNAs, yet their regulatory roles have not been well understood. Here we studied the molecular mechanisms and consequences of tsRNA-mediated regulation in Drosophila. By carrying out mRNA sequencing and ribosome profiling of S2 cells transfected with single-stranded tsRNA mimics and mocks, we show that tsRNAs recognize target mRNAs through conserved complementary sequence matching and suppress target genes by translational inhibition. Serum starvation experiments confirm tsRNAs participate in cellular starvation responses by preferential targeting the ribosomal proteins and translational initiation or elongation factors. Knock-down of AGO2 in S2 cells under normal and starved conditions reveals a dependence of the tsRNA-mediated regulation on AGO2. Our study suggests the tsRNA-mediated regulation might be crucial for the energy homeostasis and the metabolic adaptation in the cellular systems.
Project description:RNAi of signal transduction components in Drosophila S2 cells We performed RNAi knockdown experiments on 15 different signal transduction components in Drosophila S2 cells and prepared RNA libraries from the poly-adenylated fraction of the RNA. SOLiD sequencing of strand-specific, barcoded transcriptome libraries yielded expression profiles allowing us to extract common pathway signatures and indicated that Cka may function as novel regulator in Ras/MAPK signaling. ArrayExpress Release Date: 2011-07-08 Person Roles: investigator Person Last Name: Boutros Person First Name: Michael Person Email: m.boutros@dkfz.de Person Address: Deutsches Krebsforschungszentrum, Signaling and Functional Genomics (B110), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany Person Affiliation: Signaling and Functional Genomics, DKFZ Person Roles: investigator Person Last Name: Sandmann Person First Name: Thomas Person Email: t.sandmann@dkfz.de Person Address: Deutsches Krebsforschungszentrum, Signaling and Functional Genomics (B110), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany Person Affiliation: Signaling and Functional Genomics, DKFZ Person Roles: investigator Person Last Name: Horn Person First Name: Thomas Person Email: t.horn@dkfz.de Person Address: Deutsches Krebsforschungszentrum, Signaling and Functional Genomics (B110), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany Person Affiliation: Signaling and Functional Genomics, DKFZ Person Roles: submitter Person Last Name: Kerr Person First Name: Grainne Person Email: g.kerr@dkfz.de Person Address: Deutsches Krebsforschungszentrum, Signaling and Functional Genomics (B110), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany Person Affiliation: Signaling and Functional Genomics, DKFZ
Project description:Identification of the interaction partners of the protein ecdysoneless (Ecd) in Drosophila melanogaster S2 cells as well as profiling of the changes in binding for mutant, truncated Ecd del34 protein.
