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A novel role of farnesylation in targeting a mitotic checkpoint protein, human Spindly, to kinetochores.

Moudgil DK, Westcott N, Famulski JK, Patel K, Macdonald D, Hang H, Chan GK - J. Cell Biol. (2015)

Bottom Line: We showed that hSpindly is farnesylated in vivo and farnesylation is essential for its interaction with the RZZ complex and hence KT localization.FTI treatment and hSpindly knockdown displayed the same mitotic phenotypes, indicating that hSpindly is a key FTI target in mitosis.Our data show a novel role of lipidation in targeting a checkpoint protein to KTs through protein-protein interaction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.

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A proposed model of hSpindly KT localization. (A) A schematic representation of hSpindly domains and posttranslational modifications. SB, Spindly box. (B) hSpindly farnesylation induces a conformational change such that it can interact with the RZZ complex subunits Rod and Zwilch, which recruits hSpindly to KTs during mitosis. hSpindly further recruits the dynein–dynactin complex to KTs. Once all the chromosomes are aligned on the metaphase plate, hSpindly and the RZZ complex are transported from the KTs to spindle poles.
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fig8: A proposed model of hSpindly KT localization. (A) A schematic representation of hSpindly domains and posttranslational modifications. SB, Spindly box. (B) hSpindly farnesylation induces a conformational change such that it can interact with the RZZ complex subunits Rod and Zwilch, which recruits hSpindly to KTs during mitosis. hSpindly further recruits the dynein–dynactin complex to KTs. Once all the chromosomes are aligned on the metaphase plate, hSpindly and the RZZ complex are transported from the KTs to spindle poles.

Mentions: In our current structure function study, we mapped the KT localization domain of hSpindly to its C-terminal 294–605 aa (Fig. 8 A). Furthermore, our data showed that hSpindly undergoes farnesylation, which is essential for hSpindly KT localization. Substitution of the farnesylation motif with a geranylgeranylation motif does not support hSpindly KT localization. We found that FTI treatment does not interfere with KT localization of the RZZ complex, CENP-E, or CENP-F. Additionally, farnesylation plays a pivotal role in the interaction of hSpindly with the RZZ complex providing insight into lipid modification regulating checkpoint protein assembly. Our results showed that FTI treatment and hSpindly knockdown share the same phenotypes, prolonged prometaphase and metaphase with chromosome alignment defects, differing only in severity. Our analysis indicates that hSpindly is likely a key farnesylation target leading to FTI-induced mitotic defects.


A novel role of farnesylation in targeting a mitotic checkpoint protein, human Spindly, to kinetochores.

Moudgil DK, Westcott N, Famulski JK, Patel K, Macdonald D, Hang H, Chan GK - J. Cell Biol. (2015)

A proposed model of hSpindly KT localization. (A) A schematic representation of hSpindly domains and posttranslational modifications. SB, Spindly box. (B) hSpindly farnesylation induces a conformational change such that it can interact with the RZZ complex subunits Rod and Zwilch, which recruits hSpindly to KTs during mitosis. hSpindly further recruits the dynein–dynactin complex to KTs. Once all the chromosomes are aligned on the metaphase plate, hSpindly and the RZZ complex are transported from the KTs to spindle poles.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4384735&req=5

fig8: A proposed model of hSpindly KT localization. (A) A schematic representation of hSpindly domains and posttranslational modifications. SB, Spindly box. (B) hSpindly farnesylation induces a conformational change such that it can interact with the RZZ complex subunits Rod and Zwilch, which recruits hSpindly to KTs during mitosis. hSpindly further recruits the dynein–dynactin complex to KTs. Once all the chromosomes are aligned on the metaphase plate, hSpindly and the RZZ complex are transported from the KTs to spindle poles.
Mentions: In our current structure function study, we mapped the KT localization domain of hSpindly to its C-terminal 294–605 aa (Fig. 8 A). Furthermore, our data showed that hSpindly undergoes farnesylation, which is essential for hSpindly KT localization. Substitution of the farnesylation motif with a geranylgeranylation motif does not support hSpindly KT localization. We found that FTI treatment does not interfere with KT localization of the RZZ complex, CENP-E, or CENP-F. Additionally, farnesylation plays a pivotal role in the interaction of hSpindly with the RZZ complex providing insight into lipid modification regulating checkpoint protein assembly. Our results showed that FTI treatment and hSpindly knockdown share the same phenotypes, prolonged prometaphase and metaphase with chromosome alignment defects, differing only in severity. Our analysis indicates that hSpindly is likely a key farnesylation target leading to FTI-induced mitotic defects.

Bottom Line: We showed that hSpindly is farnesylated in vivo and farnesylation is essential for its interaction with the RZZ complex and hence KT localization.FTI treatment and hSpindly knockdown displayed the same mitotic phenotypes, indicating that hSpindly is a key FTI target in mitosis.Our data show a novel role of lipidation in targeting a checkpoint protein to KTs through protein-protein interaction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.

Show MeSH