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Document Type

Open Access Dissertation

Degree Name

Doctor of Philosophy (PhD)

Degree Program

Molecular and Cellular Biology

Year Degree Awarded

2018

Month Degree Awarded

September

First Advisor

Jennifer L. Ross

Subject Categories

Life Sciences

Abstract

Microtubules are polymeric protein filaments made of the monomeric alpha-beta tubulin heterodimer. They are important for physical attributes of the cell such as shape and structure, and drive essential processes such as cell division, cell migration, and active transport of metabolites. This occurs through the control of microtubules in space and over time through the lifetime of the cell. The shape and interior rearrangements are created by a diverse group of microtubule-associated proteins. When this regulation goes awry, microtubules location, stiffness, and structure are compromised, which is what is seen in diseases such as Alzheimer’s. Multiple microtubule networks can exist within a single cell under distinct spatiotemporal regulators. These networks remain poorly studied because current methods to disrupt the microtubule cytoskeleton do not easily provide rapid, local control with standard cell manipulation reagents. Here, we develop a new microtubule-disruption tool based on katanin p60 severing activity and demonstrate proof-of-principle by targeting it to kinetochores in DrosophilamelanogasterS2 cells. Specifically, we show that human katanin p60 can remove microtubule polymer mass in S2 cells and an increase in misaligned chromosomes when globally overexpressed. When targeted to the kinetochores via Mis12, we were able to recapitulate the misalignment only when using a phosphorylation-resistant mutant. Our results demonstrate that targeting an active version of katanin p60 to the kinetochore can reduce the fidelity of achieving full chromosome alignment in metaphase and could serve as a microtubule disruption tool for the future.

Available for download on Sunday, September 01, 2019

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