title: FAM29A promotes microtubule amplification via recruitment of the NEDD1-gamma-tubulin complex to the mitotic spindle.
abstract: Microtubules (MTs) are nucleated from centrosomes and chromatin. In addition, MTs can be generated from preexiting MTs in a gamma-tubulin-dependent manner in yeast, plant, and Drosophila cells, although the underlying mechanism remains unknown. Here we show the spindle-associated protein FAM29A promotes MT-dependent MT amplification and is required for efficient chromosome congression and segregation in mammalian cells. Depletion of FAM29A reduces spindle MT density. FAM29A is not involved in the nucleation of MTs from centrosomes and chromatin, but is required for a subsequent increase in MT mass in cells released from nocodazole. FAM29A interacts with the NEDD1-gamma-tubulin complex and recruits this complex to the spindle, which, in turn, promotes MT polymerization. FAM29A preferentially associates with kinetochore MTs and knockdown of FAM29A reduces the number of MTs in a kinetochore fiber, activates the spindle checkpoint, and delays the mitotic progression. Our study provides a biochemical mechanism for MT-dependent MT amplification and for the maturation of kinetochore fibers in mammalian cells.
authors.affiliation: Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhu
authors.forename: Judith A
authors.initials: JA
authors.lastname: Coppinger
authors.forename: Chang-Young
authors.initials: CY
authors.lastname: Jang
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
authors.forename: Guowei
authors.initials: G
authors.lastname: Fang
mesh_terms: D018385:Centrosome / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008938:Mitosis; D008941:Spindle Apparatus / Q000378:metabolism*; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism*
chemical_list: D002843:Chromatin; C534027:HAUS6 protein, human; D008869:Microtubule-Associated Proteins; C509189:NEDD1 protein, human; D014404:Tubulin