Project description:Measurement of capping efficiency (by 5'CAP and 5'noCAP sequencing) in male (S2) and female (Kc) Drosophila melanogaster cells upon depletion of MSL1 by dsRNA compared to the eGFP RNAi control. The measurement of capping efficiency was combined with gene expression measurement by strand specific RNA-Seq in female (Kc) Drosophila melanogaster cells
Project description:Measurement of gene expression by strand specific RNA-Seq in male (S2) Drosophila melanogaster cells upon depletion of MSL1 by dsRNA compared to the eGFP RNAi control.
Project description:The male-specific dosage compensation complex (DCC), which consists of five proteins and two non-coding roX RNAs, is necessary for the transcriptional enhancement of X-linked genes to compensate for the sex chromosome monosomy in Drosophila XY males, compared with XX females. The MSL1 and MSL2 proteins form the heterotetrameric core of DCC and are critical for the specific recruitment of the DCC to the high-affinity “entry” sites (HAS) on the X chromosome. Here we demonstrated that the N-terminal region of MSL1 is critical for its stability and functions. Amino acid deletions and substitutions in the N-terminal region of MSL1 strongly affect both interaction with roX2 RNA and DCC binding to HAS on the X chromosome. In particular, substitution of the conserved N-terminal amino-acids 3-7 in MSL1GS has an affect on dosage compensation similar to inactivation of genes encoding roX RNAs. MSL1GS binds to promoters like MSL1WT but does not co-bind with MSL2 and MSL3 to X chromosomal HAS. However, over-expression of MSL2 partially restores the functional activity of MSL1GS in dosage compensation. Thus, the interaction of MSL1 with roX RNA is critical for the efficient assembly of DCCs on HAS of the male X chromosome.
Project description:We present data using a novel method to simultaneously identify and quantify transferred male seminal proteins and the female reproductive proteome using multiplexed Tandem-Mass-Tag (TMT) isobaric labelling of the lower female reproductive tracts dissected from virgin- or recently mated- females of three species of the virilis group. We identified over 200 putative male ejaculate proteins many of which show differential abundance between species. We also identified over 2000 proteins providing the first description of the Drosophila female reproductive tract proteome outside of the melanogaster group which also shows significant divergence between species. We then assessed the utility of species-specific compared to single species query databases for protein identification and quantification.
Project description:ChIP-Seq profiles of MSL1, MSL2, MSl3, MOF, MLE, H4K16ac and RNA Polymerase II phosphorlyated on Serine 5 in Drosophila S2 cells MSL1, MSL2, MSL3, MOF, MLE, H4K16ac and RNA Polymerase II phosphorlyated on Serine 5 ChIP in Drosophila S2 cells. 1-3 biological replicates per experiment. Performed in single-read and paired-end read mode.