# Mechanics Underlying Dynein-Kinesin Bidirectional Intracellular Transport

> **NIH NIH F32** · PENNSYLVANIA STATE UNIVERSITY, THE · 2021 · $66,390

## Abstract

Project Summary
Bidirectional transport of cargo in neurons is required for their growth and function. The opposite-polarity
cytoskeletal motors, kinesin and dynein, are responsible for driving transport toward the synaptic terminals and
back to the cell center, respectively. Bidirectional transport necessarily involves a complex web of regulatory
factors from the inherent properties of the motors themselves to structural changes in the microtubule tracks
they walk on. Breakdown in the neuronal transport machinery is implicated in several neurodegenerative
diseases, including Alzheimer’s, Parkinson’s, Huntington’s and amyotrophic lateral sclerosis (ALS).
Unfortunately, the underlying mechanism of bidirectional transport is not known due to experimental limitations
that have prevented direct observation of motors working in an ensemble. A widely accepted model is “tug-of-
war”, wherein teams of kinesin and dynein pull against each other and the winning team determines the direction
of travel. However, the tug-of-war model does not account for motor-motor coordination and other possible
regulatory factors. The long-term goal of this project is to establish an in vitro method for direct observation of
motor ensembles that can be easily adapted to understand all facets of bidirectional transport. The proposed
method combines gold nanoparticle tracking via Interferometric Scattering (iSCAT) microscopy with DNA origami
to create a versatile multi-motor assay that enables precise visualization of a single motor or overall cargo
displacement during collective transport. The short-term goal of this project is to better understand the motility of
adaptor-activated mammalian dynein (DDB) as it transports cargo in a pair with a cooperative or antagonistic
partner. Aim 1 will investigate how the stepping of DDB is adapted for bidirectional movement by tracking the
motor domain as it steps with another DDB or kinesin-1. The results will shed light on why dynein works better
in a team than kinesin-1, and how dynein remains actively engaged with the cargo during kinesin-driven
transport. Aim 2 will investigate how the motility properties of the kinesin-2 and kinesin-3 families affect their
ability to compete against DDB in tug-of-war by tracking the motility of DDB-kinesin-2 and DDB-kinesin-3 pairs.
The results will provide insight into how transport is regulated by the population of kinesin families that are bound
to the cargo, and how motility properties can influence kinesin-dynein coordination. Execution of the proposed
experiments will require extensive knowledge in DNA origami design, baculovirus expression of complex
proteins, analysis of high-resolution tracking data, and construction and optimization of a high-resolution
microscope. Therefore, the overall outcome of the project will not only be significant advancement in
understanding of the connection between single motor stepping and collective transport in the cell, but also
excellent training in cut...

## Key facts

- **NIH application ID:** 10146196
- **Project number:** 5F32GM137487-02
- **Recipient organization:** PENNSYLVANIA STATE UNIVERSITY, THE
- **Principal Investigator:** Allison Gicking
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $66,390
- **Award type:** 5
- **Project period:** 2020-04-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10146196

## Citation

> US National Institutes of Health, RePORTER application 10146196, Mechanics Underlying Dynein-Kinesin Bidirectional Intracellular Transport (5F32GM137487-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10146196. Licensed CC0.

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