Enhancing Research in the STEM Disciplines at Howard University by Investigating Novel Regulation of the 26S Proteasome
Abstract
The purpose of this grant is to enhance the capacity and quality of basic, biomedical research in the function and regulation of the proteasome at Howard University. The proteasome is the major cellular complex that maintains protein homeostasis within organisms. The daily extreme stressors faced by warfighters, including emotional stress, are known to damage proteins and must elicit a proteasomal response to clear toxic proteins and maintain cellular equilibrium. Dysfunction of the proteasome has been identified as a contributing factor for numerous human conditions and diseases, including cancers and neurodegenerative disorders. Unfortunately, we do not understand the entire function of the proteasome in these conditions/diseases. In order to treat these different conditions, many of which affect our warfighters, we need to have a better understanding of all the possible positive and negative regulators of the proteasome. In this proposal we introduce a new system for studying the regulation of the proteasome. We have discovered that RNAi depletion of specific components of the Caenorhabditis elegans proteasome results in suppression of a very penetrant RNAi infertility phenotype observed upon depletion of the WEE-1.3 kinase. The overall objectives of this proposal are to fill gaps in knowledge concerning the function and regulation of the proteasome. We hypothesize that the observed suppression of the infertility phenotype is not merely a consequence of decreased proteolysis by the proteasome but rather that WEE-1.3 is a potential novel regulator of the 26S proteasome or that the proteasome is acting in a new, non-proteolytic role during the meiotic cell cycle. This hypothesis will be tested by pursuing two specific aims: (1) defining the differential roles of the various 19S proteasome subunits in the WEE-1.3 depletion phenotype; and (2) elucidating the mechanisms by which certain 19S regulatory particle (RP) subunits suppress the WEE-1.3 depletion infertility. Preliminary data shows that not every subunit within the 19S RP is capable of suppressing the wee-1.3(RNAi) infertility, and that depletion of different 19S RP subunits results in alteration in WEE-1.3 localization. This implies that the various 19S RP subunits can act functionally distinct from each other, potentially as individual entities separate from the proteasome complex. This research will lead to a significant advance in our understanding of how the proteasome is regulated. As proteasome misregulation is associated with many human diseases, understanding more about the multiple roles and functions that the proteasome plays can help guide the development of new treatment strategies for those diseases. Methodology to be utilized includes fertility assays, proteasome activity assays, live imaging microscopy, CRISPR/Cas9 endogenous genome editing, co-immunoprecipitation studies, and liquid chromatography tandem mass spectrometry (LC/MS/MS) experiments. Upon achievement of the aims, this grant will increase our understanding of the regulation of the proteasome. Additionally, this grant will increase the active participation of graduate and undergraduate underrepresented minorities (URMs) in the science and research fields. Students will be exposed to state-of-the-art endogenous genome editing technology and live imaging microscopy. By providing cutting edge, formative research experiences, these URM researchers will be more likely to continue on in STEM fields, potentially those important to the defense mission. Enabling career development opportunities and attendance at scientific meetings will help the students to have positive experiences in STEM. Finally, by providing tuition and stipend support for the students, this will alleviate some of the financial stresses faced by the students and enable them to focus more on their coursework, research, and future career development.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Feb 14, 2019
- Source ID
- W911NF1810465
Entities
People
- Anna Allen
Organizations
- Army Contracting Command
- Howard University
- Office of the Secretary of Defense