title: KLHL3Knockout Mice Reveal the Physiological Role of KLHL3 and the Pathophysiology of Pseudohypoaldosteronism Type II Caused by Mutant KLHL3.
abstract: Mutations in the with-no-lysine kinase 1 (WNK1),WNK4, kelch-like 3 (KLHL3), and cullin3 (CUL3) genes are known to cause the hereditary disease pseudohypoaldosteronism type II (PHAII). It was recently demonstrated that this results from the defective degradation of WNK1 and WNK4 by the KLHL3/CUL3 ubiquitin ligase complex. However, the other physiologicalin vivoroles of KLHL3 remain unclear. Therefore, here we generatedKLHL3-/-mice that expressed β-galactosidase (β-Gal) under the control of the endogenousKLHL3promoter. Immunoblots of β-Gal and LacZ staining revealed that KLHL3 was expressed in some organs, such as brain. However, the expression levels of WNK kinases were not increased in any of these organs other than the kidney, where WNK1 and WNK4 increased in KLHL3-/-mice but not in KLHL3+/-mice. KLHL3-/-mice also showed PHAII-like phenotypes, whereas KLHL3+/-mice did not. This clearly demonstrates that the heterozygous deletion ofKLHL3was not sufficient to cause PHAII, indicating that autosomal dominant type PHAII is caused by the dominant negative effect of mutant KLHL3. We further demonstrated that the dimerization of KLHL3 can explain this dominant negative effect. These findings could help us to further understand the physiological roles of KLHL3 and the pathophysiology of PHAII caused by mutant KLHL3.
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Emi
authors.identifier: http://orcid.org/0000-0003-2758-3309
authors.initials: E
authors.lastname: Sasaki
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Koichiro
authors.initials: K
authors.lastname: Susa
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Takayasu
authors.initials: T
authors.lastname: Mori
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Isobe
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Araki
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Inoue
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yoshizaki
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Ando
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Yutaro
authors.initials: Y
authors.lastname: Mori
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Mandai
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Moko
authors.initials: M
authors.lastname: Zeniya
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Daiei
authors.initials: D
authors.lastname: Takahashi
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Naohiro
authors.initials: N
authors.lastname: Nomura
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Tatemitsu
authors.initials: T
authors.lastname: Rai
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Uchida
authors.affiliation: Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan esohara.kid@tmd.ac.jp.
authors.forename: Eisei
authors.initials: E
authors.lastname: Sohara
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D055879:Gene Knock-In Techniques; D005799:Genes, Dominant; D007668:Kidney / Q000201:enzymology / Q000473:pathology; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008840:Microfilament Proteins / Q000172:deficiency / Q000235:genetics*; D008954:Models, Biological; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D010766:Phosphorylation; D055503:Protein Multimerization; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011546:Pseudohypoaldosteronism / Q000097:blood / Q000235:genetics* / Q000503:physiopathology*; D014018:Tissue Distribution
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C000598025:KLHL3 protein, mouse; D008840:Microfilament Proteins; D050505:Mutant Proteins; D017346:Protein Serine-Threonine Kinases
keywords: NaCl cotransporter; distal convoluted tubule; hypertension; kelch-like 3 (KLHL3); kidney; with-no-lysine kinase (WNK)