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Regulated Protein Degradation

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...

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The Proteasome

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Updated: Dec 21, 2025

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

The RING Domain of RING Finger 12 Efficiently Builds Degradative Ubiquitin Chains.

A J Middleton¹, J Zhu¹, C L Day¹

¹Department of Biochemistry, School of Biomedical Sciences, University of Otago, Dunedin 9054, New Zealand.

Journal of Molecular Biology

|May 17, 2020

Summary

This summary is machine-generated.

RNF12, a ubiquitin E3 ligase, is crucial for X-chromosome inactivation and gene regulation. Its RING domain mutations cause intellectual disability, and structural studies reveal how it interacts with enzymes for ubiquitylation.

Keywords:

RNF12 X-chromosome inactivation degradation regulation ubiquitin-conjugating

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Last Updated: Dec 21, 2025

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

In Vitro Analysis of E3 Ubiquitin Ligase Function

In Vitro Analysis of E3 Ubiquitin Ligase Function

Published on: May 14, 2021

Detection of Protein Ubiquitination

Detection of Protein Ubiquitination

Published on: August 19, 2009

Area of Science:

Biochemistry
Molecular Biology
Genetics

Background:

RNF12 is a ubiquitin E3 ligase essential for X-chromosome inactivation, transcription factor regulation, and TGF-β signaling.
Mutations in the RNF12 RING domain are linked to X-linked intellectual disability.

Purpose of the Study:

To characterize ubiquitin transfer by RNF12.
To elucidate the structural basis of RNF12 E3 ligase activity and its interaction with E2 enzymes.
To understand the impact of disease-associated mutations on RNF12 function.

Main Methods:

Ubiquitin transfer assays.
Crystal structure determination of RNF12 in complex with E2 enzymes and E2~Ub conjugates.
Comparative structural analysis with other RING domains.

Main Results:

RNF12's RING domain binds and is active with E2 enzymes that generate degradative ubiquitin chains.
Crystal structures reveal RNF12 complexed with E2 enzymes and a closed E2~Ub conformation.
A conserved loop stabilizes the active conformation of the E2~Ub conjugate.

Conclusions:

The findings provide a structural framework for understanding RNF12-mediated ubiquitylation.
This work clarifies the deleterious effects of disease-causing mutations in RNF12.
The study offers insights into the mechanism of substrate ubiquitylation by RNF12.