Directly to content
  1. Publishing |
  2. Search |
  3. Browse |
  4. Recent items rss |
  5. Open Access |
  6. Jur. Issues |
  7. DeutschClear Cookie - decide language by browser settings

RNA binding regulates TRIM25-mediated RIG-I ubiquitination

Haubrich, Kevin

[thumbnail of thesis_haubrich_2020_printed-A.pdf]
Preview
PDF, English - main document
Download (17MB) | Lizenz: Creative Commons LizenzvertragRNA binding regulates TRIM25-mediated RIG-I ubiquitination by Haubrich, Kevin underlies the terms of Creative Commons Attribution-NonCommercial-NoDerivatives 4.0

Citation of documents: Please do not cite the URL that is displayed in your browser location input, instead use the DOI, URN or the persistent URL below, as we can guarantee their long-time accessibility.

Abstract

TRIM25 is an E3 ligase of the tripartite motif protein family, that is best known for its function in innate immunity, where it activates the pattern recognition receptor RIG-I. More recently, it was identified as a putative RNA binding protein, though lacking domains with known RNA-binding potential. In this thesis, I present evidence that RNA binding is mediated by the coiled-coil (CC) and PRY/SPRY domain with possible contributions of the disordered linker connecting the domains. Using NMR spectroscopy and mutational analysis, I could map the RNA binding site on these domains. Small-angle X-ray scattering indicates that RNA-binding stabilizes an inherent, but weak interaction between these domains leading to a more rigid domain architecture possibly explaining the increase in ubiquitination activity in the presence of RNA observed by us and others. In line with that, mutants affecting RNA binding or the weak CC:PRY/SPRY interaction also reduced ubiquitination of the RIG-I caspase-activation and recruitment domains (CARDs). RNA binding in addition promotes phase-separation and association with RIG-I, as our results indicate that there is no direct protein-protein interaction between TRIM25 and RIG-I. This reconciles seemingly controversial results in recent studies and contributes to further unravel the mechanism behind the immune response activation upon viral infection.

Document type: Dissertation
Supervisor: Sachse, Prof. Dr. Carsten
Place of Publication: Heidelberg
Date of thesis defense: 15 June 2020
Date Deposited: 17 Jul 2020 09:52
Date: 2021
Faculties / Institutes: The Faculty of Bio Sciences > Dean's Office of the Faculty of Bio Sciences
Service facilities > European Molecular Biology Laboratory (EMBL)
DDC-classification: 500 Natural sciences and mathematics
570 Life sciences
Uncontrolled Keywords: TRIM25, RIG-I, RNA binding proteins, E3 ligases, innate immunity
About | FAQ | Contact | Imprint |
OA-LogoDINI certificate 2013Logo der Open-Archives-Initiative