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The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore-microtubule attachment and in maintaining the spindle assembly checkpoint.

Hauf S, Cole RW, LaTerra S, Zimmer C, Schnapp G, Walter R, Heckel A, van Meel J, Rieder CL, Peters JM - J. Cell Biol. (2003)

Bottom Line: We have identified the small molecule Hesperadin as an inhibitor of chromosome alignment and segregation.Our data imply that Hesperadin causes this phenotype by inhibiting the function of the mitotic kinase Aurora B.Hesperadin also causes cells arrested by taxol or monastrol to enter anaphase within <1 h, whereas cells in nocodazole stay arrested for 3-5 h.

View Article: PubMed Central - PubMed

Affiliation: Research Institute of Molecular Pathology, Dr. Bohr-Gasse 7, 1030 Vienna, Austria.

ABSTRACT
The proper segregation of sister chromatids in mitosis depends on bipolar attachment of all chromosomes to the mitotic spindle. We have identified the small molecule Hesperadin as an inhibitor of chromosome alignment and segregation. Our data imply that Hesperadin causes this phenotype by inhibiting the function of the mitotic kinase Aurora B. Mammalian cells treated with Hesperadin enter anaphase in the presence of numerous monooriented chromosomes, many of which may have both sister kinetochores attached to one spindle pole (syntelic attachment). Hesperadin also causes cells arrested by taxol or monastrol to enter anaphase within <1 h, whereas cells in nocodazole stay arrested for 3-5 h. Together, our data suggest that Aurora B is required to generate unattached kinetochores on monooriented chromosomes, which in turn could promote bipolar attachment as well as maintain checkpoint signaling.

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Hesperadin-treated PtK cells show a higher frequency of syntelic attachments during prometaphase than control-treated cells. (A and B) PtK1 cell treated with 500 nM Hesperadin for 3 h. Deconvolved images taken at 0.2-μm Z-interval. α-Tubulin (green), CREST (red), and DNA staining (blue). Chromosomes that are attached in a syntelic manner are marked by arrows and are enlarged in B. (C) PtK2 cells were treated with 500 nM Hesperadin or 0.1% DMSO (control) for 3 h. Deconvolution microscopy was performed as in A, and the type of attachment was determined for as many chromosomes as possible (control: 26 cells, 115 chromosomes; Hesperadin: 29 cells, 64 chromosomes; see Fig. S3, available at http://www.jcb.org/cgi/content/full/jcb.200208092/DC1). The average number of chromosomes per cell exhibiting a certain type of attachment is shown.
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fig7: Hesperadin-treated PtK cells show a higher frequency of syntelic attachments during prometaphase than control-treated cells. (A and B) PtK1 cell treated with 500 nM Hesperadin for 3 h. Deconvolved images taken at 0.2-μm Z-interval. α-Tubulin (green), CREST (red), and DNA staining (blue). Chromosomes that are attached in a syntelic manner are marked by arrows and are enlarged in B. (C) PtK2 cells were treated with 500 nM Hesperadin or 0.1% DMSO (control) for 3 h. Deconvolution microscopy was performed as in A, and the type of attachment was determined for as many chromosomes as possible (control: 26 cells, 115 chromosomes; Hesperadin: 29 cells, 64 chromosomes; see Fig. S3, available at http://www.jcb.org/cgi/content/full/jcb.200208092/DC1). The average number of chromosomes per cell exhibiting a certain type of attachment is shown.

Mentions: To determine the type of attachment of monooriented chromosomes in Hesperadin-treated cells, we performed deconvolution microscopy on PtK1 or PtK2 cells that were fixed and stained with anti-tubulin antibodies and CREST serum (Fig. 7 ; see Fig. S3, available at http://www.jcb.org/cgi/content/full/jcb.200208092/DC1). We were not able to determine the type of attachment for each chromosome of a prometaphase cell. However, we found, on average, one clearly syntelic chromosome in Hesperadin-treated cells, whereas only one in six control cells contained a clearly syntelic chromosome. Conversely, control cells contained, on average, one chromosome that could clearly be identified as monotelic, whereas this number was reduced sixfold in Hesperadin-treated cells (Fig. 7 C; Fig. S3), indicating that Aurora B function might be required to convert syntelic into monotelic attachment.


The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore-microtubule attachment and in maintaining the spindle assembly checkpoint.

Hauf S, Cole RW, LaTerra S, Zimmer C, Schnapp G, Walter R, Heckel A, van Meel J, Rieder CL, Peters JM - J. Cell Biol. (2003)

Hesperadin-treated PtK cells show a higher frequency of syntelic attachments during prometaphase than control-treated cells. (A and B) PtK1 cell treated with 500 nM Hesperadin for 3 h. Deconvolved images taken at 0.2-μm Z-interval. α-Tubulin (green), CREST (red), and DNA staining (blue). Chromosomes that are attached in a syntelic manner are marked by arrows and are enlarged in B. (C) PtK2 cells were treated with 500 nM Hesperadin or 0.1% DMSO (control) for 3 h. Deconvolution microscopy was performed as in A, and the type of attachment was determined for as many chromosomes as possible (control: 26 cells, 115 chromosomes; Hesperadin: 29 cells, 64 chromosomes; see Fig. S3, available at http://www.jcb.org/cgi/content/full/jcb.200208092/DC1). The average number of chromosomes per cell exhibiting a certain type of attachment is shown.
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fig7: Hesperadin-treated PtK cells show a higher frequency of syntelic attachments during prometaphase than control-treated cells. (A and B) PtK1 cell treated with 500 nM Hesperadin for 3 h. Deconvolved images taken at 0.2-μm Z-interval. α-Tubulin (green), CREST (red), and DNA staining (blue). Chromosomes that are attached in a syntelic manner are marked by arrows and are enlarged in B. (C) PtK2 cells were treated with 500 nM Hesperadin or 0.1% DMSO (control) for 3 h. Deconvolution microscopy was performed as in A, and the type of attachment was determined for as many chromosomes as possible (control: 26 cells, 115 chromosomes; Hesperadin: 29 cells, 64 chromosomes; see Fig. S3, available at http://www.jcb.org/cgi/content/full/jcb.200208092/DC1). The average number of chromosomes per cell exhibiting a certain type of attachment is shown.
Mentions: To determine the type of attachment of monooriented chromosomes in Hesperadin-treated cells, we performed deconvolution microscopy on PtK1 or PtK2 cells that were fixed and stained with anti-tubulin antibodies and CREST serum (Fig. 7 ; see Fig. S3, available at http://www.jcb.org/cgi/content/full/jcb.200208092/DC1). We were not able to determine the type of attachment for each chromosome of a prometaphase cell. However, we found, on average, one clearly syntelic chromosome in Hesperadin-treated cells, whereas only one in six control cells contained a clearly syntelic chromosome. Conversely, control cells contained, on average, one chromosome that could clearly be identified as monotelic, whereas this number was reduced sixfold in Hesperadin-treated cells (Fig. 7 C; Fig. S3), indicating that Aurora B function might be required to convert syntelic into monotelic attachment.

Bottom Line: We have identified the small molecule Hesperadin as an inhibitor of chromosome alignment and segregation.Our data imply that Hesperadin causes this phenotype by inhibiting the function of the mitotic kinase Aurora B.Hesperadin also causes cells arrested by taxol or monastrol to enter anaphase within <1 h, whereas cells in nocodazole stay arrested for 3-5 h.

View Article: PubMed Central - PubMed

Affiliation: Research Institute of Molecular Pathology, Dr. Bohr-Gasse 7, 1030 Vienna, Austria.

ABSTRACT
The proper segregation of sister chromatids in mitosis depends on bipolar attachment of all chromosomes to the mitotic spindle. We have identified the small molecule Hesperadin as an inhibitor of chromosome alignment and segregation. Our data imply that Hesperadin causes this phenotype by inhibiting the function of the mitotic kinase Aurora B. Mammalian cells treated with Hesperadin enter anaphase in the presence of numerous monooriented chromosomes, many of which may have both sister kinetochores attached to one spindle pole (syntelic attachment). Hesperadin also causes cells arrested by taxol or monastrol to enter anaphase within <1 h, whereas cells in nocodazole stay arrested for 3-5 h. Together, our data suggest that Aurora B is required to generate unattached kinetochores on monooriented chromosomes, which in turn could promote bipolar attachment as well as maintain checkpoint signaling.

Show MeSH
Related in: MedlinePlus