A molecular framework for control of oriented cell division in the Arabidopsis embryo
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PROVIDER: PRJNA698899 | ENA |
REPOSITORIES: ENA
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Project description:Premitotic control of cell division orientation is critical for plant development, as cell walls prevent extensive cell remodelling or migration. Whilst many divisions are proliferative and add cells to existing tissues, some divisions are formative, and generate new tissue layers or growth axes. Such formative divisions are often asymmetric in nature, producing daughters with different fates. We have previously shown that in the Arabidopsis thaliana embryo, developmental asymmetry is correlated with geometric asymmetry, creating daughter cells of unequal volume. Such divisions are generated by division planes that deviate from a default “minimal surface area” rule. Inhibition of auxin response leads to reversal to this default, yet the mechanisms underlying division plane choice in the embryo have been unclear. Here we show that auxin-dependent division plane control involves alterations in cell geometry, but not in cell polarity or nuclear position. Through transcriptome profiling, we find that auxin regulates genes controlling cell wall and cytoskeleton properties. We confirm the involvement of microtubule (MT)-binding proteins in embryo division control. Topology of both MT and Actin cytoskeleton depend on auxin response, and genetically controlled MT or Actin depolymerization in embryos leads to disruption of asymmetric divisions, including reversion to the default. Our work shows how auxin-dependent control of MT- and Actin cytoskeleton properties interacts with cell geometry to generate asymmetric divisions during the earliest steps in plant development. Transcriptional profiling of Col-0 vs bdl 8-cell Arabidopsis embryos
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Project description:Cell division is a fundamental process for multicellula organisation. Here we report a genome-scale RNAi screen in HeLa cells designed to identify human genes important for cell division. We utilized a library of endoribonuclease-prepared siRNAs (esiRNAs) for gene-silencing and used DNA content analysis to uncover genes that induced cell cycle arrest or altered ploidy upon knockdown. Rigorous validation and secondary assays were preformed to generate a nine-parameter fingerprint for each of the genes. These phenotypic signatures allowed the assignment of genes to specific functional classes by combining hierarchical clustering, cross-species analysis and proteomic data mining. We highlight the richness of our dataset by describing novel functions to genes in mitosis and cytokinesis. In particular, we identified two evolutionarily conserved transcriptional regulatory networks that govern cytokinesis in mammalian cells. Our work provides an experimental framework from which the systematic analysis of novel genes necessary for cell division in human cells can begin. An esiRNA targeting LIN54 was transfected into HCT116 cells. RNA was harvested at 24, 48, and 72 hours post-transfection for microarray analysis. Transcripts regulated at least 1.5-fold, and with a p-value<0.01 were identified as signature transcripts.
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