Date of Graduation

Summer 2023

Degree

Master of Natural and Applied Science in Biology

Department

Biology

Committee Chair

Christopher Lupfer

Abstract

Nod-like receptor family pyrin domain-containing protein 12 (NLRP12) is mainly known for its inhibitory function on NF-κB signaling in innate immune cells, and more recently, for its ability to regulate chemokine signaling and ubiquitination of the immune receptor RIG-I. Through a yeast 2-hybrid screen, the Lupfer lab previously discovered that NLRP12 interacts with other ubiquitin-associated proteins including Cullin 3 (CUL3) and RING finger protein 2 (RNF2). This research was conducted to mainly investigate the interaction between NLRP12 and CUL3 in human cells and examine the role in regulating NF-κB signaling. Previously, co-immunoprecipitation, followed by western blot analysis, and confocal microscopy confirmed the interaction in HEK293T cells. In this research, NF-κB activation was examined during the interaction. HEK293T cells that express TLR2 were co-transfected with NLRP12 and CUL3 and treated with peptidoglycan (1 µg/mL) for 0, 0.25, 0.5, 1, and 4 hours to examine NF-κB activation. Then, NF-κB activation was assessed by western blot for IκBα phosphorylation. Although NLRP12 alone suppressed NF-κB activation, the co-transfected cells did not show a significant difference from the control transfected cells. These data suggest that CUL3 negatively regulates NLRP12, preventing it from inhibiting NF-κB signaling by ubiquitinating NLRP12 itself.

Keywords

NOD-like Receptors, NLRP12, Inflammasomes, Inflammation, Autoinflammatory Disease, Ubiquitination, RING Finger Protein 2, Cullin 3

Subject Categories

Biology | Cell Biology | Laboratory and Basic Science Research | Other Cell and Developmental Biology

Copyright

© Inyeong Lee

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