Project description:mRNA expression was assessed genome-wide in brat mutant and w1118 early Drosophila embryos, to elucidate the role of BRAT in regulating mRNA stability. Four different time-points were analyzed for each genotype: 0-to-1.5 hours post-egglaying, 1.5-to-3.0 hours post-egglaying, 3.0-to-4.5 hours post-egglaying, and 4.5-to-6.0 hours post-egglaying.
Project description:BRAT-associated mRNAs and PUM-associated mRNAs were identified in early Drosophila embryos by RNA co-immunoprecipitation of the endogenous proteins using synthetic antibodies, followed by microarray analysis (RIP-Chip).
Project description:BRAT-associated mRNAs and PUM-associated mRNAs were identified in early Drosophila embryos by RNA co-immunoprecipitation of the endogenous proteins using synthetic antibodies, followed by microarray analysis (RIP-Chip). Nine RNA co-immunoprecipitations were performed. This includes 3 biological replicates each of 1) anti-BRAT RNA co-immunoprecipitations from wild-type 0-3 hour embryos, 2) anti-PUM RNA co-immunoprecipitations from wild-type 0-3 hour embryos, and 3) control antibody RNA co-immunoprecipitations from wild-type 0-3 hour embryos. BRAT samples and PUM samples were each normalized separately with the control samples, for a total of 12 processed samples (3 BRAT with 3 control normalized together, and 3 PUM with 3 control normalized together) from the 9 RNA co-immunoprecipitations.
Project description:The Drosophila TRIM-NHL protein Brain tumor (Brat) plays important roles during early embryogenesis, in cell fate decisions, during neurogenesis and in mature neurons. Brat is an RNA-binding protein and functions as translational repressor. However, which RNAs Brat regulates and how RNA-binding specificity is achieved, is unknown. Using RNA-Immunoprecipitation we identify Brat-bound mRNAs in Drosophila embryos and define a consensus binding motif. Microarrays were used to profile Brat-associated RNAs from fly embryos.
