WeightNameValue
1000 Titel
  • Translocation Biosensors – Cellular System Integrators to Dissect CRM1-Dependent Nuclear Export by Chemicogenomics
1000 Autor/in
  1. Fetz, Verena |
  2. Knauer, Shirley K. |
  3. Bier, Carolin |
  4. Von Kries, Jens Peter |
  5. Stauber, Roland |
1000 Erscheinungsjahr 2009
1000 LeibnizOpen
1000 Art der Datei
1000 Publikationstyp
  1. Artikel |
1000 Online veröffentlicht
  • 2009-07-09
1000 Erschienen in
1000 Quellenangabe
  • 9(7): 5423-5445
1000 FRL-Sammlung
1000 Copyrightjahr
  • 2009
1000 Lizenz
1000 Verlagsversion
  • http://dx.doi.org/10.3390/s90705423 |
  • https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3274152/ |
1000 Publikationsstatus
1000 Begutachtungsstatus
1000 Sprache der Publikation
1000 Abstract/Summary
  • Fluorescent protein biosensors are powerful cellular systems biology tools for dissecting the complexity of cellular processes with high spatial and temporal resolution. As regulated nucleo-cytoplasmic transport is crucial for the modulation of numerous (patho)physiological cellular responses, a detailed understanding of its molecular mechanism would open up novel options for a rational manipulation of the cell. In contrast to genetic approaches, we here established and employed high-content cellular translocation biosensors applicable for dissecting nuclear export by chemicogenomics. A431 cell lines, stably expressing a translocation biosensor composed of glutathione S-transferase, GFP and a rational combination of nuclear import and export signals, were engineered by antibiotic selection and flow cytometry sorting. Using an optimized nuclear translocation algorithm, the translocation response could be robustly quantified on the Cellomics Arrayscan® VTI platform. Subsequent to assay optimization, the assay was developed into a higher density 384-well format high-content assay and employed for the screening of the 17K ChemBioNet compound collection. This library was selected on the basis of a genetic algorithm used to identify maximum common chemical substructures in a database of annotated bioactive molecules and hence, is well-placed in the chemical space covered by bioactive compounds. Automated multiparameter data analysis combined with visual inspection allowed us to identify and to rationally discriminate true export inhibitors from false positives, which included fluorescent compounds or cytotoxic substances that dramatically affected the cellular morphology. A total of 120 potential hit compounds were selected for Cellomics Arrayscan® VTI based rescreening. The export inhibitory activity of 20 compounds effective at concentrations < 25 μM were confirmed by fluorescence microscopy in several cell lines. Interestingly, kinetic analysis allowed the identification of inhibitors capable to interfere with the export receptor CRM1-mediated nuclear export not only in an irreversible, but also in a reversible fashion. In sum, exploitation of biosensor based screening allows the identification of chemicogenomic tools applicable for dissecting nucleo-cytoplasmic transport in living cells.
1000 Sacherschließung
lokal LMB
lokal chemical biology
lokal import
lokal HIV-1 Rev
lokal nucleoporin
lokal Exportin 1/CRM1
lokal nucleocytoplasmic transport
lokal cancer
1000 Fachgruppe
  1. Biologie |
  2. Medizin |
1000 Fächerklassifikation (DDC)
1000 Liste der Beteiligten
  1. https://frl.publisso.de/adhoc/creator/RmV0eiwgVmVyZW5h|https://frl.publisso.de/adhoc/creator/S25hdWVyLCBTaGlybGV5IEsu|https://frl.publisso.de/adhoc/creator/QmllciwgQ2Fyb2xpbg==|https://frl.publisso.de/adhoc/creator/Vm9uIEtyaWVzLCBKZW5zIFBldGVy|http://orcid.org/0000-0002-1341-4523
1000 Label
1000 Förderer
  1. German Cancer Aid |
  2. Novartis-Foundation for Therapeutic Research |
  3. Peter and Traudl Engelhorn-, Kalkhof-Rose-, Wilhelm-Sander Foundation |
  4. Chemical Industry |
1000 Fördernummer
  1. FKZ: 102362
  2. -
  3. -
  4. -
1000 Förderprogramm
  1. -
  2. -
  3. -
  4. -
1000 Förderung
  1. 1000 joinedFunding-child
    1000 Förderer German Cancer Aid |
    1000 Förderprogramm -
    1000 Fördernummer FKZ: 102362
  2. 1000 joinedFunding-child
    1000 Förderer Novartis-Foundation for Therapeutic Research |
    1000 Förderprogramm -
    1000 Fördernummer -
  3. 1000 joinedFunding-child
    1000 Förderer Peter and Traudl Engelhorn-, Kalkhof-Rose-, Wilhelm-Sander Foundation |
    1000 Förderprogramm -
    1000 Fördernummer -
  4. 1000 joinedFunding-child
    1000 Förderer Chemical Industry |
    1000 Förderprogramm -
    1000 Fördernummer -
1000 Objektart article
1000 Beschrieben durch
1000 @id frl:6403244.rdf
1000 Erstellt am 2017-06-27T13:23:58.083+0200
1000 Erstellt von 25
1000 beschreibt frl:6403244
1000 Bearbeitet von 218
1000 Zuletzt bearbeitet Thu Jan 30 15:35:27 CET 2020
1000 Objekt bearb. Wed Aug 01 11:43:30 CEST 2018
1000 Vgl. frl:6403244
1000 Oai Id
  1. oai:frl.publisso.de:frl:6403244 |
1000 Sichtbarkeit Metadaten public
1000 Sichtbarkeit Daten public
1000 Gegenstand von

View source