The transmission of genetic material in each cell division requires its accurate duplication and distribution to the daughter cells. Errors in this process lead to aneuploidy, which is implicated in oncogenesis, disability and cell death. The chromosomes are segregated into two equivalent parts by a microtubule-based molecular machine, the mitotic spindle. The spindle is bipolar with each pole carrying an exact complement of chromosomes to each daughter cell. Spindle morphogenesis requires spatially controlled microtubule nucleation. The small gamma-tubulin complex is required for nucleation but is surprisingly inactive in isolated form. Our recent structural advances make a strong prediction for how the complex could be activated, which we will test by a mutational analysis. Moreover we will identify components or modifications of the complex that activate and regulate nucleation. Chromosomes attach to microtubules via kinetochores, multiprotein organelles that span from the DNA to the microtubule. This connection is carefully orchestrated and controls both the movement and organization of chromosomes to ensure proper segregation. Kinetochores perform the surprising feat of maintaining strong persistent attachments to microtubules even during the addition and removal of thousand of tubulin subunits. We will continue our reconstitution of the kinetochore from component parts to understand the molecular mechanism and regulation underlying this attachment.
The transmission of genetic material in each cell division requires its accurate duplication and distribution to the daughter cells. Errors in this process lead to aneuploidy, which is implicated in oncogenesis, disability and cell death.
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|Kim, Jae Ook; Zelter, Alex; Umbreit, Neil T et al. (2017) The Ndc80 complex bridges two Dam1 complex rings. Elife 6:|
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|Kudalkar, Emily M; Davis, Trisha N; Asbury, Charles L (2016) Preparation of Reactions for Imaging with Total Internal Reflection Fluorescence Microscopy. Cold Spring Harb Protoc 2016:pdb.prot085563|
|Kollman, Justin M; Greenberg, Charles H; Li, Sam et al. (2015) Ring closure activates yeast ?TuRC for species-specific microtubule nucleation. Nat Struct Mol Biol 22:132-7|
|Kudalkar, Emily M; Scarborough, Emily A; Umbreit, Neil T et al. (2015) Regulation of outer kinetochore Ndc80 complex-based microtubule attachments by the central kinetochore Mis12/MIND complex. Proc Natl Acad Sci U S A 112:E5583-9|
|Zelter, Alex; Bonomi, Massimiliano; Kim, Jae ook et al. (2015) The molecular architecture of the Dam1 kinetochore complex is defined by cross-linking based structural modelling. Nat Commun 6:8673|
|Tien, Jerry F; Umbreit, Neil T; Zelter, Alex et al. (2014) Kinetochore biorientation in Saccharomyces cerevisiae requires a tightly folded conformation of the Ndc80 complex. Genetics 198:1483-93|
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