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RNF213 Acts as a Molecular Switch for Cav-1 Ubiquitination and Phosphorylation in Human Cells

Jungmi Choi¹, Ryoichi Inoue¹, Yuki Masuo¹

¹Laboratory of Molecular Biosciences, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.

|June 11, 2025

Abstract:

RNF213 encodes a unique protein with AAA+ ATPase and E3 ubiquitin ligase activities that are critical for its diverse roles, which range from involvement in human vasculopathies, such as Moyamoya disease, to ubiquitination of viral and bacterial pathogens. Nevertheless, its primary functions in human signaling remain unclear due to the limited identification of direct substrates. Here, we investigated the interaction between RNF213 and caveolin-1 (Cav-1), a small scaffolding protein vital for caveolae formation and the regulation of a plethora of cellular processes. Cav-1 specifically binds within the two functional AAA+ domains of RNF213 in an ATP-dependent manner, highlighting the influence of cellular energy status on this interaction. Consequently, RNF213 ubiquitinates Cav-1 at several N-terminal lysine residues through K48 and K63 linkages, although several Moyamoya disease-associated RNF213 mutations greatly reduce this polyubiquitination. Moreover, RNF213 activity inhibits phosphorylation of a key regulatory residue of Cav-1, as RNF213 knockdown under oxidative stress markedly enhances Cav-1 Tyr14 phosphorylation and modifies nitric oxide bioavailability in endothelial cells. Collectively, our results indicate that RNF213 functions as a molecular switch modulating Cav-1 signaling based on RNF213 functionality and cellular conditions. These findings offer new insights into vascular pathogenesis and the vast signal pathways along the RNF213-Cav-1 axis.

Insights

RNF213 protein regulates caveolin-1 (Cav-1) signaling by ubiquitination, impacting vascular health. This interaction is crucial for understanding Moyamoya disease and cellular responses to stress.

Area of Science:

Biochemistry
Cell Biology
Molecular Genetics

Background:

RNF213 protein possesses AAA+ ATPase and E3 ubiquitin ligase activities, implicated in vasculopathies like Moyamoya disease.
The precise roles of RNF213 in human signaling pathways are not fully understood due to limited substrate identification.
Caveolin-1 (Cav-1) is a key scaffolding protein regulating cellular processes and caveolae formation.

Purpose of the Study:

To investigate the interaction between RNF213 and caveolin-1 (Cav-1).
To elucidate the functional consequences of RNF213-Cav-1 interaction on cellular signaling and vascular health.

Main Methods:

Investigated the binding interaction between RNF213 and Cav-1 using biochemical assays.
Assessed the ubiquitination of Cav-1 by RNF213 in an ATP-dependent manner.
Examined the effect of RNF213 on Cav-1 phosphorylation and nitric oxide bioavailability under oxidative stress.

Main Results:

Cav-1 binds to the AAA+ domains of RNF213 in an ATP-dependent manner.
RNF213 ubiquitinates Cav-1 via K48 and K63 linkages, with reduced polyubiquitination observed for Moyamoya disease-associated RNF213 mutations.
RNF213 inhibits Cav-1 phosphorylation at Tyr14, affecting nitric oxide bioavailability in endothelial cells under oxidative stress.

Conclusions:

RNF213 acts as a molecular switch modulating Cav-1 signaling based on its functionality and cellular conditions.
The RNF213-Cav-1 axis provides new insights into vascular pathogenesis, particularly Moyamoya disease.
Understanding this pathway is crucial for exploring novel therapeutic strategies for vascular disorders.

Read full analysis

Keywords:

Cav-1 MMD RNF213 caveolin-1 moyamoya disease

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