A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds.
<h4>Background</h4>Protein assemblies named kinetochores bind sister chromatids to the mitotic spindle and orchestrate sister chromatid segregation. Interference with kinetochore activity triggers a spindle checkpoint mediated arrest in mitosis, which frequently ends in cell death. We se...
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Public Library of Science (PLoS)
2010-07-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0011603&type=printable |
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| author | Emanuela Screpanti Stefano Santaguida Tam Nguyen Romano Silvestri Rick Gussio Andrea Musacchio Ernest Hamel Peter De Wulf |
| author_facet | Emanuela Screpanti Stefano Santaguida Tam Nguyen Romano Silvestri Rick Gussio Andrea Musacchio Ernest Hamel Peter De Wulf |
| author_sort | Emanuela Screpanti |
| collection | DOAJ |
| description | <h4>Background</h4>Protein assemblies named kinetochores bind sister chromatids to the mitotic spindle and orchestrate sister chromatid segregation. Interference with kinetochore activity triggers a spindle checkpoint mediated arrest in mitosis, which frequently ends in cell death. We set out to identify small compounds that inhibit kinetochore-microtubule binding for use in kinetochore-spindle interaction studies and to develop them into novel anticancer drugs.<h4>Methodology/principal findings</h4>A fluorescence microscopy-based in vitro assay was developed to screen compound libraries for molecules that prevented the binding of a recombinant human Ndc80 kinetochore complex to taxol-stabilized microtubules. An active compound was identified that acted at the microtubule level. More specifically, by localizing to the colchicine-binding site in alphabeta-tubulin the hit compound prevented the Ndc80 complex from binding to the microtubule surface. Next, structure-activity analyses distinguished active regions in the compound and led to the identification of highly potent analogs that killed cancer cells with an efficacy equaling that of established spindle drugs.<h4>Conclusions/significance</h4>The compound identified in our screen and its subsequently identified analogs represent new antitubulin chemotypes that can be synthetically developed into a novel class of antimitotic spindle drugs. In addition, they are stereochemically unique as their R- and S-isomers mimic binding of colchicine and podophyllotoxin, respectively, two antitubulin drugs that interact differently with the tubulin interface. Model-driven manipulation of our compounds promises to advance insight into how antitubulin drugs act upon tubulin. These advances in turn may lead to tailor-made colchicine site agents which would be valuable new assets to fight a variety of tumors, including those that have become resistant to the (antispindle) drugs used today. |
| format | Article |
| id | doaj-art-7ca874e015d641ce92860e9a3c6dd90f |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2010-07-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-7ca874e015d641ce92860e9a3c6dd90f2025-08-20T02:31:48ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-07-0157e1160310.1371/journal.pone.0011603A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds.Emanuela ScrepantiStefano SantaguidaTam NguyenRomano SilvestriRick GussioAndrea MusacchioErnest HamelPeter De Wulf<h4>Background</h4>Protein assemblies named kinetochores bind sister chromatids to the mitotic spindle and orchestrate sister chromatid segregation. Interference with kinetochore activity triggers a spindle checkpoint mediated arrest in mitosis, which frequently ends in cell death. We set out to identify small compounds that inhibit kinetochore-microtubule binding for use in kinetochore-spindle interaction studies and to develop them into novel anticancer drugs.<h4>Methodology/principal findings</h4>A fluorescence microscopy-based in vitro assay was developed to screen compound libraries for molecules that prevented the binding of a recombinant human Ndc80 kinetochore complex to taxol-stabilized microtubules. An active compound was identified that acted at the microtubule level. More specifically, by localizing to the colchicine-binding site in alphabeta-tubulin the hit compound prevented the Ndc80 complex from binding to the microtubule surface. Next, structure-activity analyses distinguished active regions in the compound and led to the identification of highly potent analogs that killed cancer cells with an efficacy equaling that of established spindle drugs.<h4>Conclusions/significance</h4>The compound identified in our screen and its subsequently identified analogs represent new antitubulin chemotypes that can be synthetically developed into a novel class of antimitotic spindle drugs. In addition, they are stereochemically unique as their R- and S-isomers mimic binding of colchicine and podophyllotoxin, respectively, two antitubulin drugs that interact differently with the tubulin interface. Model-driven manipulation of our compounds promises to advance insight into how antitubulin drugs act upon tubulin. These advances in turn may lead to tailor-made colchicine site agents which would be valuable new assets to fight a variety of tumors, including those that have become resistant to the (antispindle) drugs used today.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0011603&type=printable |
| spellingShingle | Emanuela Screpanti Stefano Santaguida Tam Nguyen Romano Silvestri Rick Gussio Andrea Musacchio Ernest Hamel Peter De Wulf A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds. PLoS ONE |
| title | A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds. |
| title_full | A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds. |
| title_fullStr | A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds. |
| title_full_unstemmed | A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds. |
| title_short | A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds. |
| title_sort | screen for kinetochore microtubule interaction inhibitors identifies novel antitubulin compounds |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0011603&type=printable |
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