Project description:The Arabidopsis cytochrome P450 KLUH (KLU)/CYP78A5 promotes organ growth in a non-cell autonomous manner. To identify genes regulated by KLU activity, homozygous klu-2 mutants carrying constructs for EtOH-inducible overexpression of wild-type KLU (35S::AlcR-AlcA::KLU) or of enzymatically inactive KLU protein (35S::AlcR-AlcA::KLUmut) were induced with EtOH and sampled at 90 min and 240 min after induction for gene expression changes. Keywords: Time course, genetic modification
Project description:The Clp protease complex in plants is the one with the highest complexity through all life kingdoms. It is composed of 14 nuclear- and plastid-encoded subunits which are allocated in different rings across the complex. The proteolytic ring (P-ring; ClpP3-ClpP6) and non-proteolytic ring (R-ring; ClpR1-ClpR4 and ClpP1) form the core complex playing an essential role in protein degradation and complex stabilization, respectively. The chaperones, ClpC1, ClpC2 and ClpD are involved in substrate folding and unfolding while accessory proteins, ClpT1 and ClpT2, are important to stabilize the core complex assembly. Adaptor proteins of the complex, ClpS and ClpF, recognize protease targets. Finding substrates of proteases is one of the remaining open questions in the protease field and specifically in the Clp protease. Several attempts were made analyzing the change of protein abundance in null clp mutants by proteomic analysis. However, null clp mutants are associated with massive accumulation of pleiotropic effects, as for example impaired growth and delayed in plant development, making impossible the detection of potential Clp substrates. To address this question, we design an ethanol (EtOH) inducible strategy to silence the expression of several subunits of the Clp protease in tobacco, and follow the change in protein abundance through the EtOH time-course. In this way, we distinguish between primary and secondary effects and determine time-resolved changes in proteins stability. Time-resolved proteomic analysechloroplast processes affected by the repression of Clp activity (e.g., photosynthesis and protein homeostasis).
Project description:In this study, we focused on the time-course of transcriptional changes in freshly isolated human PBMCs 4, 8, 24 and 48 h after onset of stimulation with the active vitamin D metabolite 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) compared to vehicle (0.1% EtOH).
Project description:We performed a tamoxifen-inducible, time-course ChIP-seq analysis of TBP that permits the measurement of TBP binding dynamics at target sites on a genome-wide scale in human cells.
2019-07-03 | GSE133729 | GEO
Project description:A673 Ewing's sarcoma cells: stable-knockdown and inducible-rescue time course experiments
Project description:A673 Ewing's sarcoma cells, with inducible EWS/FLI cDNA, harboring the EF-2-RNAi retrovirus, induced (or uninduced) for the indicated time period. Keywords: time course
Project description:We generated a tamoxifen-inducible skeletal muscle-specific Bmal1 knockout mouse model and performed a time-course microarray experiment to identify gene expression changes downstream of the molecular clock. We report the transcrpt expression profiles in adult gastrocnemius skeletal muscle harvested from iMS-Bmal1-/- and at 4 hour intervals for a full circadian time-course (6 time-points).
