Date of Award

Spring 2022

Thesis Type

Rollins Access Only

Degree Name

Honors Bachelor of Arts

Department

Biology

Sponsor

Dr. Jay Pieczynski

Committee Member

Dr. Pamela Brannock

Committee Member

Dr. Bobby Fokidis and Dr. James Patrone

Abstract

Kinesin motor proteins maintain the proper distribution of intracellular material by driving the transportation of specific cargo towards the positive end of microtubule tracks. While these motors have been highly studied in vitro, little is known about their in vivo function. The kinesin- 3 superfamily is highly expressed in neurons. It plays an essential role in neuronal signaling due to its ability to travel long distances by superprocessive motion. To analyze the function of kinesin- 3 motors, an in vivo study was conducted on KLP-4, the Caenorhabditis elegans homolog to human KIF13A and KIF13B. To identify interactors with KLP-4, a co-immunoprecipitation (IP) pulldown assay was performed on mixed stage C. elegans containing a 3xFLAG-tagged KLP-4 motor followed by shotgun proteomics. Of all 331 proteins, 32.6% are involved in protein production, 16.9% in cytoskeleton and membrane formation, 17.8% in protein modification and transportation, and 16.3% in cellular respiration. Together, these results suggests that KLP-4 may play a role in (1) local protein synthesis, (2) axonal growth, (3) post-translational modifications, and (4) local ATP synthesis at the ends of neurons. We also used this study to develop methods to isolate RNA from a co-IP sample, which is a highly useful protocol for future co-IP experiments. However, future research is needed to confirm the presence of ribosomal RNA in the RNA isolated from the co-IP to determine if KLP-4 plays a role in local protein synthesis in neurons.

Rights Holder

Jessika Linnemeyer

Download

Share

COinS