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Decreased KLHL3 expression is involved in the pathogenesis of pseudohypoaldosteronism type II caused by cullin 3 mutation in vivo

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Abstract

Background

Pseudohypoaldosteronism type II (PHAII) is a hereditary hypertensive disease caused by mutations in four genes: WNK1, WNK4, Kelch-like3 (KLHL3), and cullin3 (CUL3). Recently, it was revealed that CUL3–KLHL3 E3 ligase complex ubiquitinates WNK1 and WNK4, leading to their degradation, and that a common pathogenesis of PHAII is defective WNK degradation due to CUL3–KLHL3 E3 ligase complex impairment. PHAII-causing CUL3 mutations mediate exon9 skip**, producing a CUL3 protein with a 57-amino acid deletion (Δ403–459). However, the pathogenic effects of KLHL3, an adaptor protein that links WNKs with CUL3, in PHAII caused by CUL3 mutation remain unclear.

Methods

To clarify detailed pathophysiological mechanisms underlying PHAII caused by CUL3 mutation in vivo, we generated and analyzed knock-in mice carrying the same CUL3 exon9 deletion (CUL3WT/Δex9) as that reported in PHAII patients.

Results

CUL3WT/Δex9 mice exhibited a PHAII-like phenotype. Interestingly, we confirmed markedly decreased KLHL3 expression in CUL3WT/Δex9 mice by confirming the true KLHL3 band in vivo. However, the expression of other KLHL family proteins, such as KLHL2, was comparable between WT and mutant mice.

Conclusion

KLHL3 expression was decreased in CUL3WT/Δex9 mice. However, expression levels of other KLHL family proteins were comparable between the wild-type and mutant mice. These findings indicate that the decreased abundance of KLHL3 is a specific phenomenon caused by mutant CUL3 (Δexon9). Our findings would improve our understanding of the pathogenesis of PHAII caused by CUL3 mutation in vivo.

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Acknowledgements

We thank C. Iijima and M. Chiga for help in the experiments. This work is supported by the Grants-in-Aid for Scientific Research (S) from the Japanese Society for the Promotion of Science (Grant No. 25221306–00), Grants-in-Aid for Scientific Research (B) from the Japanese Society for the Promotion of Science (Grant No. 16H05314), Grant-in-Aid for Challenging Exploratory Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Author notes

  1. Sayaka Yoshida and Yuya Araki contributed equally to this work.

Authors and Affiliations

  1. Department of Nephrology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo, Tokyo, 113-8519, Japan

    Sayaka Yoshida, Yuya Araki, Takayasu Mori, Emi Sasaki, Yuri Kasagi, Kiyoshi Isobe, Koichiro Susa, Yuichi Inoue, Tomokazu Okado, Tatemitsu Rai, Shinichi Uchida & Eisei Sohara

  2. Avenir-Atip Team, INM, INSERM, University of Montpellier, 34091, Montpellier Cedex 5, France

    Pascale Bomont

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  1. Sayaka Yoshida
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Correspondence to Eisei Sohara.

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Yoshida, S., Araki, Y., Mori, T. et al. Decreased KLHL3 expression is involved in the pathogenesis of pseudohypoaldosteronism type II caused by cullin 3 mutation in vivo. Clin Exp Nephrol 22, 1251–1257 (2018). https://doi.org/10.1007/s10157-018-1593-z

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