Abstract Title

The binding mechanism of HSP90 with KEAP1 and NRF2

Start Date

16-3-2018 1:15 PM

End Date

16-3-2018 2:30 PM

Abstract Text

Background: The accumulation of oxidative damage directly connects to diseases, such as cancer and neurodegeneration, which makes studying cellular defense mechanism against oxidative stress vital. The Kelch-like ECH-associated protein1 (KEAP1)/ nuclear factor-erythroid 2 p45-related factor 2 (NRF2) antioxidant pathway and the heat shock factor 1 (HSF-1) mediated heat shock response are essential cellular cytoprotective mechanisms against oxidative stress. Under stress, Chaperone heat shock protein 90 (HSP90) disassociates from HSF-1 under stress, thus, activating HSF-1 to promotes the synthesis of cytoprotective proteins that maintain proteostasis. Also, HSP90 facilitates the post-translational modifications and the degradation of client proteins. On the other hand, KEAP1 detaches from NRF2 and promotes the production of antioxidative response elements (AREs). Unpublished data from our collaborator showed interactions of HSP90 with KEAP1 and NRF2 via yeast two-hybrid screening. These interactions could be the critical linker of the KEAP1/NRF2 pathway and heat shock response pathway. Methods: My MSc project aims to unravel the binding mechanism of HSP90 with KEAP1 and NRF2. Their interactions will be elucidated structurally in vitro and in vivo via ELISA assay, western blotting, nuclear magnetic resonance (NMR) spectroscopy, analytical ultracentrifugation, and human cell culture. Our results will establish the molecular basis of the connection between the antioxidant and heat shock response pathways via Hsp90

Results: Purification of HSP90 and KELCH domain of KEAP1 was accomplished. (project started in September 2017)

Discussion & Conclusion: None

Interdisciplinary Reflection: Interaction between HSP90 and KEAP1-NRF2 elucidates the crosstalk of two antioxidative pathways which implicates the underlying mechanism that cells use to maintain cell integrity and survive oxidative stress. Since the inhibition of HSP90-HSF1 and KEAP1-NRF2 pathways plays important roles in cancer treatment while the upregulation of them improved treatments in neurodegenerative diseases, the result will contribute to therapeutic strategy greatly.

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Mar 16th, 1:15 PM Mar 16th, 2:30 PM

The binding mechanism of HSP90 with KEAP1 and NRF2

Background: The accumulation of oxidative damage directly connects to diseases, such as cancer and neurodegeneration, which makes studying cellular defense mechanism against oxidative stress vital. The Kelch-like ECH-associated protein1 (KEAP1)/ nuclear factor-erythroid 2 p45-related factor 2 (NRF2) antioxidant pathway and the heat shock factor 1 (HSF-1) mediated heat shock response are essential cellular cytoprotective mechanisms against oxidative stress. Under stress, Chaperone heat shock protein 90 (HSP90) disassociates from HSF-1 under stress, thus, activating HSF-1 to promotes the synthesis of cytoprotective proteins that maintain proteostasis. Also, HSP90 facilitates the post-translational modifications and the degradation of client proteins. On the other hand, KEAP1 detaches from NRF2 and promotes the production of antioxidative response elements (AREs). Unpublished data from our collaborator showed interactions of HSP90 with KEAP1 and NRF2 via yeast two-hybrid screening. These interactions could be the critical linker of the KEAP1/NRF2 pathway and heat shock response pathway. Methods: My MSc project aims to unravel the binding mechanism of HSP90 with KEAP1 and NRF2. Their interactions will be elucidated structurally in vitro and in vivo via ELISA assay, western blotting, nuclear magnetic resonance (NMR) spectroscopy, analytical ultracentrifugation, and human cell culture. Our results will establish the molecular basis of the connection between the antioxidant and heat shock response pathways via Hsp90

Results: Purification of HSP90 and KELCH domain of KEAP1 was accomplished. (project started in September 2017)

Discussion & Conclusion: None

Interdisciplinary Reflection: Interaction between HSP90 and KEAP1-NRF2 elucidates the crosstalk of two antioxidative pathways which implicates the underlying mechanism that cells use to maintain cell integrity and survive oxidative stress. Since the inhibition of HSP90-HSF1 and KEAP1-NRF2 pathways plays important roles in cancer treatment while the upregulation of them improved treatments in neurodegenerative diseases, the result will contribute to therapeutic strategy greatly.