title: A novel role of farnesylation in targeting a mitotic checkpoint protein, human Spindly, to kinetochores.
abstract: 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.
authors.affiliation: Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.
authors.forename: Devinderjit K
authors.initials: DK
authors.lastname: Moudgil
authors.affiliation: Laboratory of Chemical Biology and Microbial Pathogenesis, Rockefeller University, New York, NY 10065.
authors.forename: Nathan
authors.initials: N
authors.lastname: Westcott
authors.affiliation: Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.
authors.forename: Jakub K
authors.initials: JK
authors.lastname: Famulski
authors.affiliation: Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.
authors.forename: Kinjal
authors.initials: K
authors.lastname: Patel
authors.affiliation: Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.
authors.forename: Dawn
authors.initials: D
authors.lastname: Macdonald
authors.affiliation: Laboratory of Chemical Biology and Microbial Pathogenesis, Rockefeller University, New York, NY 10065.
authors.forename: Howard
authors.initials: H
authors.lastname: Hang
authors.affiliation: Department of Oncology, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2 gkc@ualberta.ca.
authors.forename: Gordon K T
authors.initials: GK
authors.lastname: Chan
mesh_terms: D002352:Carrier Proteins / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D020090:Chromosome Segregation / Q000235:genetics*; D051231:Farnesyltranstransferase / Q000037:antagonists & inhibitors; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D059566:M Phase Cell Cycle Checkpoints / Q000235:genetics*; D061986:MCF-7 Cells; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics; D009154:Mutation; D017368:Protein Prenylation / Q000235:genetics*; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D034741:RNA, Small Interfering; D017384:Sequence Deletion; D008941:Spindle Apparatus / Q000235:genetics
chemical_list: D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; C489309:KNTC1 protein, human; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; C549402:SPDL1 protein, human; C488261:ZW10 protein, human; C068688:centromere protein E; C094772:centromere protein F; D051231:Farnesyltranstransferase
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