KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathway
Abstract Background Studies reported that kelch-like protein 3 (KLHL3)-Cullin3(CUL3) E3 ligase ubiquitinated with-no-lysine kinase 4 (WNK4). Impaired WNK4 ubiquitination plays a key role in Familial hyperkalemic hypertension (FHHt, also called pseudohypoaldosteronism type II) which results from over...
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| Format: | Article |
| Language: | English |
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BMC
2023-07-01
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| Series: | BMC Nephrology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12882-023-03257-4 |
| _version_ | 1827895303929331712 |
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| author | Siqi Ying Qin Guo Chong Zhang |
| author_facet | Siqi Ying Qin Guo Chong Zhang |
| author_sort | Siqi Ying |
| collection | DOAJ |
| description | Abstract Background Studies reported that kelch-like protein 3 (KLHL3)-Cullin3(CUL3) E3 ligase ubiquitinated with-no-lysine kinase 4 (WNK4). Impaired WNK4 ubiquitination plays a key role in Familial hyperkalemic hypertension (FHHt, also called pseudohypoaldosteronism type II) which results from overaction of thiazide-sensitive sodium chloride cotransport (NCC). In addition, researchers have also found that dietary potassium deficiency activates NCC along the renal distal convoluted tubule (DCT). However, the underlying mechanism remains unclear about the relationship between potassium and WNK4. Methods In the present study, we conducted in vitro and in vivo experiments to confirm that KLHL3-dependent WNK4 degradation is affected by potassium through the neddylation and autophagy pathway. In vitro, the WNK4 and KLHL3 plasmids were cotransfected into HEK293 cell lines by lipofectamine 2000, and then incubated with different potassium concentrations (1mmol/L and 10mmol/L) for 24 h, and further treated with MLN4924 or the autophagy inhibitor or both of MLN4924 and the autophagy inhibitor for another 24 h respectively. In vivo, we created mice that were fed with low or high potassium diets and then were injected MLN4924 in the experimental groups. The expression of WNK4, pWNK4, KLHL3, NEDD8, LC3 ,and P62 was detected by western blotting in vitro and vivo experiments. Results We found that the abundance and phosphorylation of WNK4 increase when neddylation is inhibited both in vitro and vivo. Furthermore, the abundance of pWNK4, WNK4, NEDD8, and KLHL3 was increased in the low potassium (LK) group. Inhibiting autophagy can ameliorate the effect of potassium on the abundance and activity of WNK4 to some extent. Conclusion These findings suggest a complex regulation of potassium in the degradation of WNK4. Low potassium can activate WNK4, which may be related to neddylation and autophagy, but the mechanism needs to be further studied. |
| first_indexed | 2024-03-12T22:19:06Z |
| format | Article |
| id | doaj.art-b778260dc5d540f593e6349977f53ac9 |
| institution | Directory Open Access Journal |
| issn | 1471-2369 |
| language | English |
| last_indexed | 2024-03-12T22:19:06Z |
| publishDate | 2023-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Nephrology |
| spelling | doaj.art-b778260dc5d540f593e6349977f53ac92023-07-23T11:08:59ZengBMCBMC Nephrology1471-23692023-07-012411810.1186/s12882-023-03257-4KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathwaySiqi Ying0Qin Guo1Chong Zhang2Department of Nephrology, Jing’an District Center Hospital of Shanghai, Fudan UniversityDepartment of Nephrology, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of MedicineDepartment of Nephrology, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of MedicineAbstract Background Studies reported that kelch-like protein 3 (KLHL3)-Cullin3(CUL3) E3 ligase ubiquitinated with-no-lysine kinase 4 (WNK4). Impaired WNK4 ubiquitination plays a key role in Familial hyperkalemic hypertension (FHHt, also called pseudohypoaldosteronism type II) which results from overaction of thiazide-sensitive sodium chloride cotransport (NCC). In addition, researchers have also found that dietary potassium deficiency activates NCC along the renal distal convoluted tubule (DCT). However, the underlying mechanism remains unclear about the relationship between potassium and WNK4. Methods In the present study, we conducted in vitro and in vivo experiments to confirm that KLHL3-dependent WNK4 degradation is affected by potassium through the neddylation and autophagy pathway. In vitro, the WNK4 and KLHL3 plasmids were cotransfected into HEK293 cell lines by lipofectamine 2000, and then incubated with different potassium concentrations (1mmol/L and 10mmol/L) for 24 h, and further treated with MLN4924 or the autophagy inhibitor or both of MLN4924 and the autophagy inhibitor for another 24 h respectively. In vivo, we created mice that were fed with low or high potassium diets and then were injected MLN4924 in the experimental groups. The expression of WNK4, pWNK4, KLHL3, NEDD8, LC3 ,and P62 was detected by western blotting in vitro and vivo experiments. Results We found that the abundance and phosphorylation of WNK4 increase when neddylation is inhibited both in vitro and vivo. Furthermore, the abundance of pWNK4, WNK4, NEDD8, and KLHL3 was increased in the low potassium (LK) group. Inhibiting autophagy can ameliorate the effect of potassium on the abundance and activity of WNK4 to some extent. Conclusion These findings suggest a complex regulation of potassium in the degradation of WNK4. Low potassium can activate WNK4, which may be related to neddylation and autophagy, but the mechanism needs to be further studied.https://doi.org/10.1186/s12882-023-03257-4KLHL3CUL3WNK4PotassiumNeddylationAutophagy |
| spellingShingle | Siqi Ying Qin Guo Chong Zhang KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathway BMC Nephrology KLHL3 CUL3 WNK4 Potassium Neddylation Autophagy |
| title | KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathway |
| title_full | KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathway |
| title_fullStr | KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathway |
| title_full_unstemmed | KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathway |
| title_short | KLHL3-dependent WNK4 degradation affected by potassium through the neddylation and autophagy pathway |
| title_sort | klhl3 dependent wnk4 degradation affected by potassium through the neddylation and autophagy pathway |
| topic | KLHL3 CUL3 WNK4 Potassium Neddylation Autophagy |
| url | https://doi.org/10.1186/s12882-023-03257-4 |
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