Understanding Cellular Dynamics: Investigating a Yeast-Two Hybrid Protein-Protein Interaction, RIOK3:CSE1L; also by Investigating Trogocytosis of CD4+ T Cells from RhoG-/- Mice

Author

Brittnee N. Crane, University of Montana, MissoulaFollow

Year of Award

2024

Document Type

Thesis

Degree Type

Master of Science (MS)

Degree Name

Cellular, Molecular, and Microbial Biology (Microbiology and Immunology)

Department or School/College

DBS, CMMB

Committee Chair

Dr. Scott Wetzel

Committee Co-chair

Dr. Steve Lodmell

Commitee Members

Dr. Scott Wetzel, Dr. Steve Lodmell, and Dr. Bruce Bowler

Keywords

RIOK3, CSE1L, Immune pathways, Trogocytosis, RhoG Knockout, RhoG Independent Trogocytosis

Publisher

University of Montana

Subject Categories

Animal Experimentation and Research | Biology | Cell Biology | Cellular and Molecular Physiology | Immunology and Infectious Disease | Laboratory and Basic Science Research | Microbiology | Virology

Abstract

Rift Valley fever virus (RVFV) is a negative-sense virus that was first described in 1930 when it was isolated from sheep during an outbreak. In the last 25 years there has been a spread in the geographical range into the Middle East. The most susceptible animals are cattle, goats, sheep, and camels. The most overt symptom of livestock infection is that infected pregnant animals experience spontaneous abortion. Humans are susceptible as well and symptoms range from mild febrile illness to liver necrosis, blindness, hemorrhagic fever, and death. An important player in the cellular innate response to RVFV infections is the human protein, RIOK3. RIOK3 is a serine/threonine-protein kinase involved in the regulation of type I interferon (IFN)-dependent immune responses, which happens to play a crucial role in the innate responses to many viruses. RIOK3 has proven necessary in mounting the IFN antiviral response in epithelial cells post-infection with RVFV. Because RIOK3 is critical for responding to RVFV infection and other viruses of clinical or agricultural significance, it is of compelling interest to know how RIOK3 performs its antiviral functions. To gain insight into RIOK3’s function, we carried out a yeast-two hybrid (Y2H) experiment to discover novel protein-protein interactions with RIOK3. We identified the human protein Xpo2 as a candidate interactor in the Y2H screen and then subjected it to siRNA knockdown to assess its role in the cellular antiviral response.

Trogocytosis, dynamic process of cell-to-cell communication, significantly influences immune response, development and disease. A newly published study has even shown red blood cells (RBCs) as capable of conducting trogocytosis, which has significant clinical impacts. Researchers continue to explore the multifaceted nature, seeing novel therapeutic insights. In this study, we aimed to enhance trogocytosis potential in non-polarized TCR-transgenic CD4⁺ T cells. We evaluated various commercial antibody methods for their impact on trogocytosis. Previous work by Martinez-Martin et al. highlighted RhoG’s critical role in CD4⁺ T cell trogocytosis, revealing that TC21 was essential for TCR internalization with the mechanism dependent on RhoG. Our study challenges the conventional view that RhoG is indispensable for trogocytosis. By investigating alternative pathways, we contribute to a deeper understanding of cellular interactions and immune responses. Further research into the molecular mechanisms underlying trogocytosis promises valuable insight for therapeutic strategies and disease management

Recommended Citation

Crane, Brittnee N., "Understanding Cellular Dynamics: Investigating a Yeast-Two Hybrid Protein-Protein Interaction, RIOK3:CSE1L; also by Investigating Trogocytosis of CD4+ T Cells from RhoG-/- Mice" (2024). Graduate Student Theses, Dissertations, & Professional Papers. 12358.
https://scholarworks.umt.edu/etd/12358