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Please use this identifier to cite or link to this item: http://arks.princeton.edu/ark:/88435/dsp01bg257j430
Title: Microtubule formation via the microtubule-associated protein HURP
Authors: Valdez, Venecia Alexandria
Advisors: Petry, Sabine
Contributors: Molecular Biology Department
Keywords: DLGAP5
HURP
Microtubule
Subjects: Molecular biology
Biochemistry
Issue Date: 2024
Publisher: Princeton, NJ : Princeton University
Abstract: During cell division, the microtubule-based spindle is assembled to align and segregate chromosomes. A vital process of spindle assembly is the nucleation of the microtubules that form the spindle structure. Spindle microtubule nucleation and spindle function is facilitated by many microtubule-associated proteins (MAPs). Here, I review the current spindle assembly field and investigate the roles of MAPs, HURP and Eg5, that have been previously implicated in spindle microtubule formation.I find that HURP is necessary for branching microtubule nucleation and promotes the general formation of microtubules in meiotic Xenopus laevis egg extract. Conversely, my findings suggest that the motor protein, Eg5, does not affect branching microtubule nucleation. Additionally, I find that TPX2 and HURP work synergistically to promote branching microtubule nucleation in egg extract. I provide a cryo-EM structure of the HURP microtubule binding domain bound to the microtubule, revealing that HURP may serve to stabilize the microtubule lattice. In support of this, I find that HURP reduces incidences of catastrophe and increases rescue in a dynamic microtubule assay in vitro. Furthermore, I provide preliminary data to suggest that the microtubule binding domain of HURP alone is sufficient to promote microtubule nucleation in the egg extract. I combine extract, in vitro, and structural approaches to decipher the function of HURP in microtubule formation. This function may imply a role for HURP in the formation of kinetochore fibers, where HURP is known to localize and contribute to chromosome alignment, thus further highlighting the importance of MAPs in spindle assembly.
URI: http://arks.princeton.edu/ark:/88435/dsp01bg257j430
Type of Material: Academic dissertations (Ph.D.)
Language: en
Appears in Collections:Molecular Biology

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