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Global Analysis of Translational Reprogramming during Flagellar Regeneration Reveals the Role of Sphingolipid Metabolism in Ciliogenesis and Ciliopathies

ABSTRACT: Global Analysis of Translational Reprogramming during Flagellar Regeneration Reveals the Role of Sphingolipid Metabolism in Ciliogenesis and Ciliopathies

PROVIDER: PRJNA630259 | ENA |

REPOSITORIES: ENA

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Global Analysis of Translational Reprogramming during Flagellar Regeneration Reveals the Role of Sphingolipid Metabolism in Ciliogenesis and Ciliopathies

Project description:Protein synthesis is essential for cilium assembly, but little is known about translational reprogramming that occurs during ciliogenesis. Using ribosome footprinting, we perform global translatome profiling on Chlamydomonas during flagellar regeneration.

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Role for KDM3A in ciliogenesis

Project description:Cilia are dynamic antennae that sense the extracellular environment adjusting their length to maintain homeostasis. Mutant mouse models for the lysine demethylase KDM3A (JMJD1A, JMHD2A) share phenotypic features with human ciliopathies, including obesity and metabolic syndrome. Here we show that cilia of Kdm3a mutants are unstable with dysregulated length ranges accumulating intraflagellar transport proteins (IFTs) at the ciliary tip. RNA sequencing and mass-spectrometry identified actin cytoskeleton as the most miss-regulated feature during the ciliary cycle of KDM3A null cells and revealed that IFT81 contains Nε-methylated lysines in vivo to which recombinant KDM3A binds without subsequent demethylation. Mutations in these IFT81 methyl-lysine residues however stabilize KDM3A null cilia and potentiate ciliogenesis surpassing wild type cells; a synergism phenocopied by wild type cultures through the simultaneous destabilization of the actin cytoskeleton when over-expressing IFT81 lysine-mutants. Our work reveals that ciliogenesis requires the coordinated release of cytoskeletal constrains and the presence of intraflagellar transport proteins which KDM3A integrates aided by post-translationally modifiable lysine residues in IFT81.

2018-07-11 | PXD004334 | Pride

A ciliopathy complex restrains centriolar length and builds distal appendages to initiate ciliogenesis.

Project description:Mitosis segregates into each daughter cell two centrioles, the older of which is uniquely capable of generating a cilium. How this older centriole, called the mother centriole, initiates ciliogenesis remains poorly understood. We have identified an evolutionarily conserved complex comprised of CEP90, OFD1, MNR and FOPNL. Human mutations in CEP90, MNR and OFD1 cause ciliopathies. Super-resolution microscopy revealed that this complex forms a ring at the distal centriole. Centrioles of cells lacking MNR or CEP90 failed to assemble distal appendages and cannot generate cilia. In addition to the centrioles, complex members localized to centriolar satellites, proteinaceous granules surrounding the centrioles. Disruption of satellites did not affect distal appendage assembly, indicating that the centriolar pool is required for ciliogenesis. Consistent with an essential role in ciliogenesis, mice lacking MNR or CEP90 did not assemble cilia, did not survive beyond embryonic day 9.5, and did not transduce Hedgehog signals. In addition to ciliogenesis, MNR, but not CEP90, restrained centriolar length. MNR recruited both OFD1, required for centriolar length control, and CEP90, which recruits CEP83 to root distal appendages. Thus, an evolutionarily conserved ciliopathy-associated complex functions at the distal centriole to control centriole length and assemble distal appendages.

2021-06-15 | PXD022372 | Pride

Lysinoalanine crosslinking is a conserved post-translational modification in the spirochete flagellar hookLysinoalanine crosslinking is a conserved post-translational modification in the spirochete flagellar hook

Project description:In this project, we investigated whether diverse spirochete species contain lysinoalanine cross-linkages between conserved residues in their flagellar hook proteins (FlgE). We examined the following species: Treponema denticola¸ Borreliella burgdorferi, Treponema phagedenis, Treponema pallidum, Brachyspira hyodysenteriae, Leptospira interrogans. For each bacterial species, we examined a variety of different sample types, including recombinantly produced FlgE, polyhooks purified from flik knock-out strains, and wild-type periplasmic filaments. Overall, our data suggests that lysinoalanine crosslinking in a conserved FlgE post-translational modification in spirochetes and is required for the unique motility of these organisms. For more information, please see the following manuscript: https://doi.org/10.1101/2023.06.13.544825.

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Project description:Treating Saccharomyces cerevisiae cells with myriocin reduces the intracellular free pool of most amino acids. We find that transcriptional changes within the first 6 hours of drug treatment seem to have little impact on free pool changes. Rather the changes are driven early by post-translational events including impaired amino acid transporter activity, due to a reduced rate of sphingolipid biosynthesis, and an increased rate of endocytosis of at least some amino acid transporters, particularly the major methionine transporter Mup1.

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MG1655 Flagellar Regulators RNA-seq

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The polar flagellar transcriptional regulatory network in Vibrio campbellii deviates from known Vibrio species

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Transcriptional organization and regulation of the Pseudomonas putida flagellar system

Project description:The entire set of flagellar structural components and flagellar-specific transcriptional regulators, as well as much of the core chemotaxis machinery, is encoded into a >70 kbp cluster in Pseudomonas putida KT2440 genome. We have performed RNA-seq of the wild-type strain in order to identify operon boundaries and promoters location in this cluster.

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sphingo-1-Identification of Sphingolipid regulated genes in cold stress response. Sphingolipids regulated genes.

Project description:rs07-05_sphingolipids-cold - sphingo-1 - The cold choc response seems to be partly triggered by Sphingolipid species. To date no gene response as been associated to sphingolipid signaling pathway in plant. Our aim is to identify among the cold induced genes the ones regulated by sphingolipids and to try to define a sphingolipid pathway specific group of genes. - 7ml of 5 days-old cells suspensions were incubated in presence of different sphingolipid pathway inhibitors, 30 min to 2 hours depending in the coumpound (all were resuspended in DMSO and control were done with DMSO). Then a 30 min cold choc was applied before cells were harvested and frozen in cold nitrogen. RNA were then extracted. FB1 and DMS were from Alexis , Myr from Cayman, TSP from matreya. 6 dye-swap - treated vs untreated comparison

2012-03-27 | E-GEOD-36659 | biostudies-arrayexpress

MG1655 Flagellar Regulators RNA-seq

Project description:This experiment contains RNA-seq data for a motile derivative of MG1655 and isogenic strains with deletions of each flagellar regulator: flhD, flhC, and fliA. All strains were grown at 37 degrees C in LB. After RNA purification, ribosomal RNA was removed using RiboZero and a library was made of remaining total RNA.

2014-08-01 | E-MTAB-2802 | biostudies-arrayexpress

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