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Publication Date

2023-05-15

First Advisor

Nathan D. Derr

Document Type

Honors Project

Degree Name

Bachelor of Arts

Department

Biochemistry

Keywords

molecular motor proteins, dynein, DNA origami, dynein ensembles, intracellular cargo transport, biophysics, single molecule microscopy

Abstract

The microtubule associated motor protein cytoplasmic dynein is central to eukaryotic, intracellular cargo transport. In cells, these molecular motors work as teams in which multiple motors are connected to a single cargo. Despite the physiological importance of these motor ensembles and a growing knowledge base surrounding individual motor mechanisms, the biophysics of motor cooperation on a single cargo is poorly understood. Through the use of programmable DNA origami “chassis” structures and total internal reflectance microscopy, we are able to monitor the motility of specially designed ensembles in which the motor number and arrangement as well as the placement and degree of resistive force are controlled. This study seeks to make use of these methods to discern if, and how, the applied forces of disparate motors are structurally, and possibly synergistically, integrated through independent connections to a resistive load. Specifically, I hope to compare the motive forces of ensembles arranged with shared and independent cargo attachment sites and investigate the effects of motor number and resistive load placement on force integration and transmission through cargo structures. We hypothesize that individual motors are better able to share a load when independently attached to a cargo because such configurations may reduce negative interference between motors and allow for the direct application of each motor’s individual force on the resistive load. This thesis focuses on the development and optimization of methods for use to investigate the emergent motility of synthetic motor ensembles in these arrangements.

Rights

©2023 Hana Hieshima. Access limited to the Smith College community and other researchers while on campus. Smith College community members also may access from off-campus using a Smith College log-in. Other off-campus researchers may request a copy through Interlibrary Loan for personal use.

Language

English

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