Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2
Abstract Loss-of-function mutations in the human gene encoding the neuron-specific Ca2+ channel CaV2.1 are linked to the neurological disease episodic ataxia type 2 (EA2), as well as neurodevelopmental disorders such as developmental delay and developmental epileptic encephalopathy. Disease-associat...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-11-01
|
| Series: | Cell Communication and Signaling |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12964-024-01960-9 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1846147623102185472 |
|---|---|
| author | Ssu-Ju Fu Kai-Min Cheng Cheng-Tsung Hsiao Ya-Ching Fang Chung-Jiuan Jeng Chih-Yung Tang |
| author_facet | Ssu-Ju Fu Kai-Min Cheng Cheng-Tsung Hsiao Ya-Ching Fang Chung-Jiuan Jeng Chih-Yung Tang |
| author_sort | Ssu-Ju Fu |
| collection | DOAJ |
| description | Abstract Loss-of-function mutations in the human gene encoding the neuron-specific Ca2+ channel CaV2.1 are linked to the neurological disease episodic ataxia type 2 (EA2), as well as neurodevelopmental disorders such as developmental delay and developmental epileptic encephalopathy. Disease-associated CaV2.1 mutants may exhibit defective proteostasis and promote endoplasmic reticulum (ER)-associated degradation of their wild-type (WT) counterpart in a dominant-negative manner. The E3 ubiquitin ligase RNF138 was previously shown to mediate EA2-related aberrant degradation of CaV2.1 at the ER. Herein we aimed to elucidate the ER proteostasis mechanism of CaV2.1. The peptidyl-prolyl cis/trans isomerase, NIMA-interacting 1 (Pin1) was identified as a novel neuronal CaV2.1 binding partner that promoted polyubiquitination and proteasomal degradation of CaV2.1. Suppression of endogenous Pin1 level with either shRNA knockdown or the Pin1 inhibitor all-trans retinoic acid (ATRA) enhanced endogenous CaV2.1 protein level in neurons, and attenuated ER-associated degradation of CaV2.1 WT and EA2-causing mutants. Detailed mutation analyses suggested that Pin1 interacted with specific phosphorylated serine/threonine-proline motifs in the intracellular II-III loop and the distal carboxy-terminal region of human CaV2.1. We further generated Pin1-insensitive CaV2.1 constructs and demonstrated that, during ER quality control, Pin1 served as an upstream regulator of CaV2.1 polyubiquitination and degradation by RNF138. Pin1 regulation was required for the dominant-negative effect of EA2 missense mutants, but not nonsense mutants, on CaV2.1 WT protein expression. Our data are consistent with the idea that CaV2.1 proteostasis at the ER, as well as dominant-negative suppression of disease-causing loss-of-function mutants on CaV2.1 WT, entail both Pin1/RNF138-dependent and -independent mechanisms. |
| format | Article |
| id | doaj-art-c101447efafa40999fb74b6482cc9c59 |
| institution | Kabale University |
| issn | 1478-811X |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj-art-c101447efafa40999fb74b6482cc9c592024-12-01T12:37:11ZengBMCCell Communication and Signaling1478-811X2024-11-0122112610.1186/s12964-024-01960-9Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2Ssu-Ju Fu0Kai-Min Cheng1Cheng-Tsung Hsiao2Ya-Ching Fang3Chung-Jiuan Jeng4Chih-Yung Tang5Department of Physiology, College of Medicine, National Taiwan UniversityDepartment of Physiology, College of Medicine, National Taiwan UniversityDepartment of Physiology, College of Medicine, National Taiwan UniversityInstitute of Anatomy and Cell Biology, College of Medicine, National Yang Ming Chiao Tung UniversityInstitute of Anatomy and Cell Biology, College of Medicine, National Yang Ming Chiao Tung UniversityDepartment of Physiology, College of Medicine, National Taiwan UniversityAbstract Loss-of-function mutations in the human gene encoding the neuron-specific Ca2+ channel CaV2.1 are linked to the neurological disease episodic ataxia type 2 (EA2), as well as neurodevelopmental disorders such as developmental delay and developmental epileptic encephalopathy. Disease-associated CaV2.1 mutants may exhibit defective proteostasis and promote endoplasmic reticulum (ER)-associated degradation of their wild-type (WT) counterpart in a dominant-negative manner. The E3 ubiquitin ligase RNF138 was previously shown to mediate EA2-related aberrant degradation of CaV2.1 at the ER. Herein we aimed to elucidate the ER proteostasis mechanism of CaV2.1. The peptidyl-prolyl cis/trans isomerase, NIMA-interacting 1 (Pin1) was identified as a novel neuronal CaV2.1 binding partner that promoted polyubiquitination and proteasomal degradation of CaV2.1. Suppression of endogenous Pin1 level with either shRNA knockdown or the Pin1 inhibitor all-trans retinoic acid (ATRA) enhanced endogenous CaV2.1 protein level in neurons, and attenuated ER-associated degradation of CaV2.1 WT and EA2-causing mutants. Detailed mutation analyses suggested that Pin1 interacted with specific phosphorylated serine/threonine-proline motifs in the intracellular II-III loop and the distal carboxy-terminal region of human CaV2.1. We further generated Pin1-insensitive CaV2.1 constructs and demonstrated that, during ER quality control, Pin1 served as an upstream regulator of CaV2.1 polyubiquitination and degradation by RNF138. Pin1 regulation was required for the dominant-negative effect of EA2 missense mutants, but not nonsense mutants, on CaV2.1 WT protein expression. Our data are consistent with the idea that CaV2.1 proteostasis at the ER, as well as dominant-negative suppression of disease-causing loss-of-function mutants on CaV2.1 WT, entail both Pin1/RNF138-dependent and -independent mechanisms.https://doi.org/10.1186/s12964-024-01960-9Peptidyl-prolyl cis/trans isomeraseE3 ubiquitin ligaseProteasomal degradationProteostasisER quality controlChannelopathy |
| spellingShingle | Ssu-Ju Fu Kai-Min Cheng Cheng-Tsung Hsiao Ya-Ching Fang Chung-Jiuan Jeng Chih-Yung Tang Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2 Cell Communication and Signaling Peptidyl-prolyl cis/trans isomerase E3 ubiquitin ligase Proteasomal degradation Proteostasis ER quality control Channelopathy |
| title | Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2 |
| title_full | Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2 |
| title_fullStr | Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2 |
| title_full_unstemmed | Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2 |
| title_short | Pin1 promotes human CaV2.1 channel polyubiquitination by RNF138: pathophysiological implication for episodic ataxia type 2 |
| title_sort | pin1 promotes human cav2 1 channel polyubiquitination by rnf138 pathophysiological implication for episodic ataxia type 2 |
| topic | Peptidyl-prolyl cis/trans isomerase E3 ubiquitin ligase Proteasomal degradation Proteostasis ER quality control Channelopathy |
| url | https://doi.org/10.1186/s12964-024-01960-9 |
| work_keys_str_mv | AT ssujufu pin1promoteshumancav21channelpolyubiquitinationbyrnf138pathophysiologicalimplicationforepisodicataxiatype2 AT kaimincheng pin1promoteshumancav21channelpolyubiquitinationbyrnf138pathophysiologicalimplicationforepisodicataxiatype2 AT chengtsunghsiao pin1promoteshumancav21channelpolyubiquitinationbyrnf138pathophysiologicalimplicationforepisodicataxiatype2 AT yachingfang pin1promoteshumancav21channelpolyubiquitinationbyrnf138pathophysiologicalimplicationforepisodicataxiatype2 AT chungjiuanjeng pin1promoteshumancav21channelpolyubiquitinationbyrnf138pathophysiologicalimplicationforepisodicataxiatype2 AT chihyungtang pin1promoteshumancav21channelpolyubiquitinationbyrnf138pathophysiologicalimplicationforepisodicataxiatype2 |