Accurate chromosome segregation is required for propagation of all cells. Errors in this process are implicated in oncogenesis, birth defects and cell death. Crucial to proper partitioning of the chromosomes during cell division is the establishment, arrangement and then breakdown of a bipolar spindle.
The aim of this grant is to further understand the structure, assembly and dynamics of the proteins that create and control the organization of the spindle in the model organism Saccharomyces cerevisiae. The microtubule organizing center in yeast is the spindle pole body (SPB). The SPB is a dynamic structure that undergoes remodeling in a cell-cycle specific manner. We have identified proteins involved in the remodeling process and will characterize them. We have provided a detailed architectural description of the SPB and will continue our structural analysis. The SPB nucleates and organizes microtubules. We will perform a set of experiments designed to distinguish between two current models for nucleation. Finally, we have found that kinetochores can biorient without being attached to the plus-end of microtubules. New insights into biorientation will be obtained from determining the role of kinetochores, motors and other spindle proteins in the formation of the bipolar spindle when plus-end attachment is not involved. Because many proteins and spindle features are conserved from yeast to human, our studies will inform models of how eukaryotic cells correctly distribute their genetic material.
|Kim, Jae Ook; Zelter, Alex; Umbreit, Neil T et al. (2017) The Ndc80 complex bridges two Dam1 complex rings. Elife 6:|
|Kudalkar, Emily M; Deng, Yi; Davis, Trisha N et al. (2016) Coverslip Cleaning and Functionalization for Total Internal Reflection Fluorescence Microscopy. Cold Spring Harb Protoc 2016:pdb.prot085548|
|Kudalkar, Emily M; Davis, Trisha N; Asbury, Charles L (2016) Single-Molecule Total Internal Reflection Fluorescence Microscopy. Cold Spring Harb Protoc 2016:pdb.top077800|
|Hsia, Yang; Bale, Jacob B; Gonen, Shane et al. (2016) Design of a hyperstable 60-subunit protein dodecahedron. [corrected]. Nature 535:136-9|
|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|
|Umbreit, Neil T; Miller, Matthew P; Tien, Jerry F et al. (2014) Kinetochores require oligomerization of Dam1 complex to maintain microtubule attachments against tension and promote biorientation. Nat Commun 5:4951|
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