Analysis of the role of KLHL3/Cullin3 in PHAII

• Project/Area Number: 25461243
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Kidney internal medicine
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Ohta Akihito 東京医科歯科大学, 医歯(薬)学総合研究科, 非常勤講師 (00510356)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 遺伝性高血圧 / WNKキナーゼ / 偽性低アルドステロン症II型 / ユビキチン化 / 遺伝子改変マウス / ノックインマウス / Akt / リン酸化 / NaCl共輸送体(NCC) / NaCl共輸送体（NCC） / インスリン / バゾプレシン / 質量分析 / PHAII / ユビキチン

Outline of Final Research Achievements

Recently, Kelch-like 3 (KLHL3) and Cullin3 (Cul3) were identified as novel causative genes for the hereditary hypertensive disease, pseudohypoaldopsteronism type II (PHAII). To elucidate the relationship between KLHL3/Cul3 and the WNK signaling pathway,　we analyzed protein overexpression system in cultured cells and generated mutant Cul3 knock-in mice. As a result, we found that KLHL3 was associated with the ubiquitination of WNK1/4, and that mutant KLHL3 interfered with WNK ubiquitination resulting in the hyper-activation of the WNK signaling pathway, leading to the development of hypertension. In the mutant Cul3 knock-in mice, decreased expression of Cullin3 was observed, whereas PHAII-like phenotype was not replicated. This result suggested that mere heterozygous deletion of Cul3 was not sufficient for the onset of PHAII and that the mutant Cul3 protein with the deletion of exon9 might have a dominant-negative function.

Research Products

• [Journal Article] Generation and analysis of knock-in mice carrying pseudohypoaldosteronism type II-causing mutations in the cullin 3 gene. (2015)
  • Author(s): Araki Y, Rai T, Sohara E, Mori T, Inoue Y, Isobe K, Kikuchi E, Ohta A, Sasaki S, Uchida S.
  • Journal Title: Biol Open Volume: 4 Issue: 11 Pages: 1509-1517
  • DOI: 10.1242/bio.013276
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Impaired degradation of WNK by Akt and PKA phosphorylation of KLHL3. (2015)
  • Author(s): Yoshizaki Y, Mori Y, Tsuzaki Y, Mori T, Nomura N, Wakabayashi M, Takahashi D, Zeniya M, Kikuchi E, Araki Y, Ando F, Isobe K, Nishida H, Ohta A, Susa K, Inoue Y, Chiga M, Rai T, Sasaki S, Uchida S, Sohara E.
  • Journal Title: Biochem Biophys Res Commun. Volume: 467 Issue: 2 Pages: 229-234
  • DOI: 10.1016/j.bbrc.2015.09.184
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Acidic motif in WNKs are crucial components for the binding to KLHL3 (2013)
  • Author(s): Akihito Ohta, Shinichi Uchida, Sei Sasaki, Dario R. Alessi
  • Organizer: American society of Nephrology annual meeting
  • Place of Presentation: アトランタ、GA、USA