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Analysis of the cilia membrane proteome by cmAPEX

ABSTRACT: APEX was targeted to the primary cilium of mouse IMCD cells using a 5HT6-APEX-GFP fusion construct. Proximity labling was performed using incubation with biotinphenol and H2O2. For details, please see the methods section of the accompanying paper.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Kidney

DISEASE(S): Mixed Disorder As Reaction To Stress

SUBMITTER: Markus Rinschen

LAB HEAD: Bernhard Schermer

PROVIDER: PXD005201 | Pride | 2017-07-18

REPOSITORIES: Pride

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			Action	DRS
	20160303_UC_ColID_97_Priyanka_Kohli_Benzing_0139_160229PrKoS01.raw	Raw
	20160303_UC_ColID_97_Priyanka_Kohli_Benzing_0139_160229PrKoS02.raw	Raw
	20160321_UC_ColID_97_Priyanka_Kohli_Benzing_0139_160307PrKoS01.raw	Raw
	20160321_UC_ColID_97_Priyanka_Kohli_Benzing_0139_160307PrKoS02.raw	Raw
	20160412_UC_ColID_129_Benzing_Priyanka_Kohli_0139_160410PrKo_S01.raw	Raw

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Publications

The ciliary membrane-associated proteome reveals actin-binding proteins as key components of cilia.

Kohli Priyanka P Höhne Martin M Jüngst Christian C Bertsch Sabine S Ebert Lena K LK Schauss Astrid C AC Benzing Thomas T Rinschen Markus M MM Schermer Bernhard B

EMBO reports 20170714 9

Primary cilia are sensory, antennae-like organelles present on the surface of many cell types. They have been involved in a variety of diseases collectively termed ciliopathies. As cilia are essential regulators of cell signaling, the composition of the ciliary membrane needs to be strictly regulated. To understand regulatory processes at the ciliary membrane, we report the targeting of a genetically engineered enzyme specifically to the ciliary membrane to allow biotinylation and identification ...[more]

PMID: 28710093

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MicroRNA (miRNA) expression of IMCD tubule suspension

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Primary cilia control translation and the cell cycle in medulloblastoma

Project description:The primary cilium, a signaling organelle projecting from the surface of a cell, controls cellular physiology and behavior. The presence or absence of primary cilia is a distinctive feature of a given tumor type; however, whether and how the primary cilium contributes to tumorigenesis is unknown for most tumors. Medulloblastoma (MB) is a common pediatric brain cancer comprising four groups: SHH, WNT, group 3 (G3), and group 4 (G4). From 111 cases of MB, we show that primary cilia are abundant in SHH and WNT MBs but rare in G3 and G4 MBs. Using WNT and G3 MB mouse models, we show that primary cilia promote WNT MB by facilitating translation of mRNA encoding β-catenin, a major oncoprotein driving WNT MB, whereas cilium loss promotes G3 MB by disrupting cell cycle control and destabilizing the genome. Our findings reveal tumor type–specific ciliary functions and underlying molecular mechanisms. Moreover, we expand the function of primary cilia to translation control and reveal a molecular mechanism by which cilia regulate cell cycle progression, providing new frameworks for studying cilia function in normal and pathologic conditions.

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Loss of primary cilia drives switching from Hedgehog to Ras/MAPK pathway

Project description:Basal cell carcinomas (BCCs) rely on Hedgehog (HH) pathway growth signal amplification by the microtubule-based organelle, the primary cilium. Despite naïve tumors responsiveness to Smoothened inhibitors (Smoi), resistance in advanced tumors remains frequent. While the resistant BCCs usually maintain HH pathway activation, squamous cell carcinomas with Ras/MAPK pathway activation also arise, with the molecular basis of tumor type and pathway selection still obscure. Here we identify the primary cilium as a critical determinant controlling tumor pathway switching. Strikingly, Smoi-resistant BCCs possess an increased mutational load in ciliome genes, resulting in reduced primary cilia and HH pathway activation compared to naive or Gorlin patient BCCs. Gene set enrichment analysis of resistant BCCs with a low HH pathway signature reveals increased Ras/MAPK pathway activation. Tissue analysis confirms an inverse relationship between primary cilia presence and Ras/MAPK activation, and primary cilia removal in BCCs potentiates Ras/MAPK pathway activation. Moreover, activating Ras in HH-responsive cell lines confers resistance to both canonical (vismodegib) and non-canonical (aPKC and MRTF inhibitors) HH pathway inhibitors, while conferring sensitivity to MAPK inhibitors. Our results provide new insights into BCC treatment and identify the primary cilium as an important lineage gatekeeper, preventing HH to Ras/MAPK pathway switching.

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Toxoplasma GRA protein identification

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2018-11-02 | PXD009604 | Pride

Piwi is a key regulator of both somatic and germline stem cells in the Drosophila testis

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Transcription profiling of rat inner medullary collecting duct

Project description:Identification of gene expressed in the enriched inner medullary collecting duct cells in rat. Experiment Overall Design: Rat inner medullary collecting duct (IMCD) cells were isolated from 7 male Sprage-Dawley rats by collagenase and hyaluronidase digestion and follow by low speed centrifugation. The non-IMCD cells were collected by centrifugation of supernatant of enriched IMCD samples. Experiment Overall Design: Total RNA about 3 ug were used per microarray (Rat 230 2.0 Genechip array). Experiment Overall Design: The experiments were repeat 3 times (3 pairs of IMCD VS non-IMCD)

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The lymph node transcriptome of unicentric and idiopathic multicentric Castleman disease

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