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Important finding in the quest for 'plant caspases': VPEs are essential for the hypersensitive cell death and react with caspase probes. VPEs also share structural homologies with caspases, but their subcellular location and substrate specificity is clearly different.
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Proteolysis of RuBisCo during victorin-induced programmed cell death can be inhibited with various protease inhibitors, including the caspase inhibitor VAD-cmk. Purification of the corresponding proteases uncovered that subtilase-like serine proteases can also react with the caspase probe VAD-cmk, illustrating the low specificity of this small molecule.
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Nagata N., Min Y.K., Nakano T., Asami T., and Yoshida S. Treatment of dark-grown Arabidopsis thaliana with a brassinosteroid-biosynthesis inhibitor, brassinazole, induces some characteristics of light-grown plants. Planta 211 (2000) 781-790
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Nagata N., Asami T., and Yoshida S. Brassinazole, an inhibitor of brassinosteroid biosysnthesis, inhibits development of secondary xylem in cress plants (Lepidium sativum). Plant Cell Physiol 42 (2001) 1006-1011
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Good example of using chemical interference to study processes in non-model organisms. Together with earlier work [12-14], these data illustrate the specificity and applicability of Brz.
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Wang Z.Y., Nakano T., Gendron J., He J., Chen M., Vafeados D., Yang Y., Fujioka S., Yoshida S., Asami T., and Chory J. Nuclear-localized BZR1 mediates brassinosteroid-induced growth and feedback suppression of brassinosteroid biosynthesis. Dev Cell 2 (2002) 505-513
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BZR1 is a transcriptional repressor with dual roles in brassinosteroid homeostasis and growth responses
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BZR1 was originally identified from a screen for mutants that were insensitive for brassinazole [16], a clear example of how chemical tools can be used to address biological questions.
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He J.X., Gendron J.M., Sun Y., Gampala S.S.L., Gendron N., Sun C.Q., and Wang Z.Y. BZR1 is a transcriptional repressor with dual roles in brassinosteroid homeostasis and growth responses. Science 307 (2005) 1634-1638. BZR1 was originally identified from a screen for mutants that were insensitive for brassinazole [16], a clear example of how chemical tools can be used to address biological questions.
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One of the few real chemical genetic screens on plants. Of the 10 000 diverse small molecules, 30 interfere in auxin signalling and four were further studied. This study illustrates the potential of nonbiased chemical genetic screens.
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Dai X., Hayashi K., Nozaki H., Cheng Y., and Zhao Y. Genetic and chemical analysis of the action mechanisms of sirtinol in Arabidopsis. Proc Natl Acad Sci USA 102 (2005) 3129-3134. Remarkable discoveries are described concerning the deceiving action of sirtinol, uncovered by collaboration with organic chemists, who produced sirtinol derivatives that act as an auxin. This work, together with the identification of sirtinol [27] and the identification of Sir1 [28], illustrates the difficulties that can be encountered by chemical genetic approaches.
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One of the earliest true chemical genetic screens of large chemical libraries to find modulators that disturb vacuolar trafficking in yeast. Only a few of the 14 bioactive compounds had similar effects in plants.
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The power of chemical genomics to study the link between endomembrane system components and the gravitropic response
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One of the few true forward chemical genetic screens using large diverse libraries is described. A simple assay, based on rotation of plates with seedlings, was used to identify small molecules that interfere in the gravitropic response.
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Surpin M., Rojas-Pierce M., Carter C., Hicks G.R., Vasquez J., and Raikhel N.V. The power of chemical genomics to study the link between endomembrane system components and the gravitropic response. Proc Natl Acad Sci USA 102 (2005) 4902-4907. One of the few true forward chemical genetic screens using large diverse libraries is described. A simple assay, based on rotation of plates with seedlings, was used to identify small molecules that interfere in the gravitropic response.
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Activity profiling of papain-like cysteine proteases in plants
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A first step towards guided reverse chemical genetics in plants: a E64-derived probe displays the activities of at least six different papain-like cysteine proteases.
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Van der Hoorn R.A.L., Leeuwenburgh M.A., Bogyo M., Joosten M.H.A.J., and Peck S. Activity profiling of papain-like cysteine proteases in plants. Plant Physiol 135 (2004) 1170-1178. A first step towards guided reverse chemical genetics in plants: a E64-derived probe displays the activities of at least six different papain-like cysteine proteases.
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