A novel role of farnesylation in targeting a mitotic checkpoint protein, human Spindly, to kinetochores.
A novel role of farnesylation in targeting a mitotic checkpoint protein, human Spindly, to kinetochores.
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DOI：
10.1083/jcb.201412085
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发表时间：
2015-03-30
期刊：
影响因子：
--
通讯作者：
Chan GK
中科院分区：
文献类型：
--
作者：
Moudgil DK;Westcott N;Famulski JK;Patel K;Macdonald D;Hang H;Chan GK
有丝分裂检查点蛋白Spindly在体内被法尼基化，这种修饰是其与RZZ复合物相互作用及其定位于着丝粒所必需的。有丝分裂检查点蛋白的动粒定位对于它们在有丝分裂期间的功能是必不可少的。hSpindly KT定位依赖于RZZ复合物，并且hSpindly在有丝分裂期间将动力蛋白-动力肌动蛋白复合物募集到KT，但hSpindly KT募集的机制尚不清楚。通过结构域定位研究，我们表征了hSpindly的KT定位结构域，并发现其在C-末端半胱氨酸残基处经历法尼基化。hSpindly的N-末端293个残基被定位为KT。使用法尼基转移酶抑制剂（FTI）抑制法尼基化废除了hSpindly KT定位，而不影响RZZ复合物、CENP-E和CENP-F KT定位。我们发现，hSpindly是法尼基化在体内和法尼基化是必不可少的与RZZ复合物的相互作用，因此KT定位。FTI处理和hSpindly敲低显示出相同的有丝分裂表型，表明hSpindly是有丝分裂中的关键FTI靶标。我们的数据显示了脂化在通过蛋白质-蛋白质相互作用将检查点蛋白靶向KT中的新作用。
The mitotic checkpoint protein Spindly is farnesylated in vivo and this modification is required for its interaction with the RZZ complex and its localization to kinetochores. Kinetochore (KT) localization of mitotic checkpoint proteins is essential for their function during mitosis. hSpindly KT localization is dependent on the RZZ complex and hSpindly recruits the dynein–dynactin complex to KTs during mitosis, but the mechanism of hSpindly KT recruitment is unknown. Through domain-mapping studies we characterized the KT localization domain of hSpindly and discovered it undergoes farnesylation at the C-terminal cysteine residue. The N-terminal 293 residues of hSpindly are dispensable for its KT localization. Inhibition of farnesylation using a farnesyl transferase inhibitor (FTI) abrogated hSpindly KT localization without affecting RZZ complex, CENP-E, and CENP-F KT localization. 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.
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影响因子：
--
作者：
Charron G;Tsou LK;Maguire W;Yount JS;Hang HC
通讯作者：
Hang HC
DOI：
10.1073/pnas.92.7.2984
发表时间：
1995-03-28
影响因子：
11.1
作者：
BHATTACHARYA, S;CHEN, L;POWERS, S
通讯作者：
POWERS, S
影响因子：
3.3
作者：
Barisic M;Sohm B;Mikolcevic P;Wandke C;Rauch V;Ringer T;Hess M;Bonn G;Geley S
通讯作者：
Geley S
DOI：
10.1038/nrm3255
发表时间：
2011-12-22
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
影响因子：
4.8
作者：
Crespo, NC;Ohkanda, J;Sebti, SM
通讯作者：
Sebti, SM