Interaction Between Two E3 ligases, NEDD8ylated Cullin and HHARI

RBR (RING1-in between RING-RING2) is a special type of E3 ubiquitin ligase containing three zinc-binding RING (Really Interesting New Gene) domains, while adopting mechanisms of HECT (Homologous to E6-AP Carboxyl Terminus) for substrate ubiquitination. Most well known RBRs include Parkin and HOIP, which are associated with Parkinson’s disease and innate immune deficiency. However, it is not well known how the RBR proteins gain activity, as they are known to be autoinhibited. Here I show that a specific F430A, E431A, E503A triple mutation of RBR protein HHARI (Human homologue of Ariadne) and its interaction with NEDD8ylated cullin RING ligase can both boost its activity and stabilize complex formation. Analytical size-exclusion chromatography, autoubiquitination, and electron microscopy reveal consistent behavior for this triple-mutant. Future structure-based studies will help elucidate the mechanism of the unsolved mystery of RBR activation and its interaction with NEDD8ylated cullin RING ligases.

Name thesis:degree_name

Master of Science (MS)

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Biomedical Sciences

Year dc:date.available

2016

Author dc:creator

• Baek, Kheewoong

Contributors dc:contributor

• Brenda A. Schulman, Ph.D.

• ubiquitin
• Chemicals and Drugs
• Enzymes and Coenzymes
• Medical Biochemistry
• Medical Immunology
• Medical Sciences
• Medicine and Health Sciences

Repository record dc:identifier

https://dc.uthsc.edu/dissertations/392

OAI identifier oai:identifier

oai:dc.uthsc.edu:dissertations-1383