The cullin 4A/B-DDB1-cereblon E3 ubiquitin ligase complex mediates the degradation of CLC-1 chloride channels

Yi An Chen, Yi Jheng Peng, Meng Chun Hu, Jing Jia Huang, Yun Chia Chien, June Tai Wu, Tsung-Yu Chen, Chih Yung Tang

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Voltage-gated CLC-1 chloride channels play a critical role in controlling the membrane excitability of skeletal muscles. Mutations in human CLC-1 channels have been linked to the hereditary muscle disorder myotonia congenita. We have previously demonstrated that disease-associated CLC-1 A531V mutant protein may fail to pass the endoplasmic reticulum quality control system and display enhanced protein degradation as well as defective membrane trafficking. Currently the molecular basis of protein degradation for CLC-1 channels is virtually unknown. Here we aim to identify the E3 ubiquitin ligase of CLC-1 channels. The protein abundance of CLC-1 was notably enhanced in the presence of MLN4924, a specific inhibitor of cullin-RING E3 ligases. Subsequent investigation with dominant-negative constructs against specific subtypes of cullin-RING E3 ligases suggested that CLC-1 seemed to serve as the substrate for cullin 4A (CUL4A) and 4B (CUL4B). Biochemical examinations further indicated that CUL4A/B, damage-specific DNA binding protein 1 (DDB1), and cereblon (CRBN) appeared to co-exist in the same protein complex with CLC-1. Moreover, suppression of CUL4A/B E3 ligase activity significantly enhanced the functional expression of the A531V mutant. Our data are consistent with the idea that the CUL4A/B-DDB1-CRBN complex catalyses the polyubiquitination and thus controls the degradation of CLC-1 channels.

Original languageEnglish (US)
Article number10667
JournalScientific Reports
Volume5
DOIs
StatePublished - May 29 2015

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Cullin Proteins
Chloride Channels
Ubiquitin-Protein Ligases
DNA-Binding Proteins
Proteolysis
Myotonia Congenita
Membranes
Muscular Diseases
Mutant Proteins
Endoplasmic Reticulum
Quality Control
CLC-1 channel
Skeletal Muscle
Proteins
Mutation

ASJC Scopus subject areas

  • General

Cite this

Chen, Y. A., Peng, Y. J., Hu, M. C., Huang, J. J., Chien, Y. C., Wu, J. T., ... Tang, C. Y. (2015). The cullin 4A/B-DDB1-cereblon E3 ubiquitin ligase complex mediates the degradation of CLC-1 chloride channels. Scientific Reports, 5, [10667]. https://doi.org/10.1038/srep10667

The cullin 4A/B-DDB1-cereblon E3 ubiquitin ligase complex mediates the degradation of CLC-1 chloride channels. / Chen, Yi An; Peng, Yi Jheng; Hu, Meng Chun; Huang, Jing Jia; Chien, Yun Chia; Wu, June Tai; Chen, Tsung-Yu; Tang, Chih Yung.

In: Scientific Reports, Vol. 5, 10667, 29.05.2015.

Research output: Contribution to journalArticle

Chen, Yi An ; Peng, Yi Jheng ; Hu, Meng Chun ; Huang, Jing Jia ; Chien, Yun Chia ; Wu, June Tai ; Chen, Tsung-Yu ; Tang, Chih Yung. / The cullin 4A/B-DDB1-cereblon E3 ubiquitin ligase complex mediates the degradation of CLC-1 chloride channels. In: Scientific Reports. 2015 ; Vol. 5.
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abstract = "Voltage-gated CLC-1 chloride channels play a critical role in controlling the membrane excitability of skeletal muscles. Mutations in human CLC-1 channels have been linked to the hereditary muscle disorder myotonia congenita. We have previously demonstrated that disease-associated CLC-1 A531V mutant protein may fail to pass the endoplasmic reticulum quality control system and display enhanced protein degradation as well as defective membrane trafficking. Currently the molecular basis of protein degradation for CLC-1 channels is virtually unknown. Here we aim to identify the E3 ubiquitin ligase of CLC-1 channels. The protein abundance of CLC-1 was notably enhanced in the presence of MLN4924, a specific inhibitor of cullin-RING E3 ligases. Subsequent investigation with dominant-negative constructs against specific subtypes of cullin-RING E3 ligases suggested that CLC-1 seemed to serve as the substrate for cullin 4A (CUL4A) and 4B (CUL4B). Biochemical examinations further indicated that CUL4A/B, damage-specific DNA binding protein 1 (DDB1), and cereblon (CRBN) appeared to co-exist in the same protein complex with CLC-1. Moreover, suppression of CUL4A/B E3 ligase activity significantly enhanced the functional expression of the A531V mutant. Our data are consistent with the idea that the CUL4A/B-DDB1-CRBN complex catalyses the polyubiquitination and thus controls the degradation of CLC-1 channels.",
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