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Characterization of inhibitors of glucocorticoid receptor nuclear translocation: a model of cytoplasmic dynein-mediated cargo transport

, : Characterization of inhibitors of glucocorticoid receptor nuclear translocation: a model of cytoplasmic dynein-mediated cargo transport. Assay and Drug Development Technologies 10(1): 46-60

Agonist-induced glucocorticoid receptor [GR] transport from the cytoplasm to the nucleus was used as a model to identify dynein-mediated cargo transport inhibitors. Cell-based screening of the library of pharmacologically active compound (LOPAC)-1280 collection identified several small molecules that stalled the agonist-induced transport of GR-green fluorescent protein (GFP) in a concentration-dependent manner. Fluorescent images of microtubule organization, nuclear DNA staining, expression of GR-GFP, and its subcellular distribution were inspected and quantified by image analysis to evaluate the impact of compounds on cell morphology, toxicity, and GR transport. Given the complexity of the multi-protein complex involved in dynein-mediated cargo transport and the variety of potential mechanisms for interruption of that process, we therefore developed and validated a panel of biochemical assays to investigate some of the more likely intracellular target(s) of the GR transport inhibitors. Although the apomorphine enantiomers exhibited the most potency toward the ATPase activities of cytoplasmic dynein, myosin, and the heat-shock proteins (HSPs), their apparent lack of specificity made them unattractive for further study in our quest. Other molecules appeared to be nonspecific inhibitors that targeted reactive cysteines of proteins. Ideally, specific retrograde transport inhibitors would either target dynein itself or one of the other important proteins associated with the transport process. Although the hits from the cell-based screen of the LOPAC-1280 collection did not exhibit this desired profile, this screening platform provided a promising phenotypic system for the discovery of dynein/HSP modulators.


PMID: 21919741

DOI: 10.1089/adt.2010.0367

Other references

Williamson, R.M.; Oxender, D.L., 1990: Sequence and structural similarities between the leucine-specific binding protein and leucyl-tRNA synthetase of Escherichia coli. A role for the leucyl-tRNA synthetase (EC has been established for regulating the transport of leucine across the inner membrane of Escherichia coli by the leucine, isoleucine, valine (LIV-I) transport system. This transport system is med...

Maddison, G.E., 1965: Increasing Difficulties In The Diagnosis Of Tuberculosis. Canadian Journal of Public Health 56: 103-104

Tartari, A.; Casnati, E.; Fernandez, J.E.; Felsteiner, J.; Baraldi, C., 1994: Photon backscattering tissue characterization by energy dispersive spectroscopy evaluations. Techniques for in vivo tissue characterization based on scattered photons have usually been confined to evaluating coherent and Compton peaks. However, information can also be obtained from the energy analysis of the Compton scattered distribution...

Edwards, Lynne, 1998: Limit values. Fenger, J [Editor], Hertel, O [Editor], Palmgren, F [Editor] Environmental Pollution, Vol 1; Urban air pollution: European aspects 419-432

Auge R.M.; Schekel K.A.; Wample R.L., 1986: Osmotic adjustment in leaves of vesicular arbuscular mycorrhizal and nonmycorrhizal rose rosa hybrida cultivar samantha plants in response to drought stress. Osmotic adjustment in Rosa hybrida L. cv. Samantha was characterized by the pressure-volume approach in drought-acclimated and unacclimated plants brought to the same level of drought strain, as assayed by stomatal closure. Plants were colonized b... J.B.; Hoffman L.J.; Menn J.J.; Casida J.E., 1971: Dyfonate metabolism studies part 2 metabolic pathway of o ethyl s phenylethyl phosphonodithioate in rats. Pesticide Biochemistry & Physiology 1(3-4): 356-365

Cloutier Y., 1989: Growth and freezing tolerance of alfalfa cell cultures. Plant Physiology (Rockville) 89(4 SUPPL): 26

Butini, S.; Nikolic, K.; Kassel, S.; Brückmann, H.; Filipic, S.; Agbaba, D.; Gemma, S.; Brogi, S.; Brindisi, M.; Campiani, G.; Stark, H., 2016: Polypharmacology of dopamine receptor ligands. Most neurological diseases have a multifactorial nature and the number of molecular mechanisms discovered as underpinning these diseases is continuously evolving. The old concept of developing selective agents for a single target does not fit with...

Muszynski, B., 1975: Blood cells of diagnostic importance in bovine lymphatic leukosis. Medycyna weterynaryjna: 31 (7) 431-432

Pascual, Mercedes, 1993: Diffusion-induced chaos in a spatial predator-prey system. A continuous predator-prey model in which both species diffuse along a spatial gradient is shown to exhibit temporal chaos at a fixed point in space. The model incorporates a nonlinear functional response of the predator and a logistic growth of t...