Gyuyeong Rah1, Hwayeon Cha1, Joohee Kim1, Jieun Song2, Hyunho Kim3, Yun Kyu Oh4, Curie Ahn5, Minyong Kang6, Jongmin Kim1, Kyung Hyun Yoo1, Min Jung Kim1, Hyuk Wan Ko2, Je Yeong Ko7, Jong Hoon Park7. 1. Department of Biological Science, Sookmyung Women's University, Seoul, Korea. 2. Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea. 3. Center for Medical Innovation, Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea. 4. Department of Internal Medicine, Seoul National University Boramae Medical Center, Seoul, Korea. 5. Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea. 6. Department of Urology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. 7. Department of Biological Science, Sookmyung Women's University, Seoul, Korea jeyeong@sookmyung.ac.kr parkjh@sookmyung.ac.kr.
Abstract
BACKGROUND: Ciliogenesis-associated kinase 1 (CILK1) is a ciliary gene that localizes in primary cilia and regulates ciliary transport. Mutations in CILK1 cause various ciliopathies. However, the pathogenesis of CILK1-deficient kidney disease is unknown. METHODS: To examine whether CILK1 deficiency causes PKD accompanied by abnormal cilia, we generated mice with deletion of Cilk1 in cells of the renal collecting duct. A yeast two-hybrid system and coimmunoprecipitation (co-IP) were used to identify a novel regulator, kinesin light chain-3 (KLC3), of ciliary trafficking and cyst progression in the Cilk1-deficient model. Immunocytochemistry and co-IP were used to examine the effect of KLC3 on ciliary trafficking of the IFT-B complex and EGFR. We evaluated the effects of these genes on ciliary trafficking and cyst progression by modulating CILK1 and KLC3 expression levels. RESULTS: CILK1 deficiency leads to PKD accompanied by abnormal ciliary trafficking. KLC3 interacts with CILK1 at cilia bases and is increased in cyst-lining cells of CILK1-deficient mice. KLC3 overexpression promotes ciliary recruitment of IFT-B and EGFR in the CILK1 deficiency condition, which contributes to the ciliary defect in cystogenesis. Reduction in KLC3 rescued the ciliary defects and inhibited cyst progression caused by CILK1 deficiency. CONCLUSIONS: Our findings suggest that CILK1 deficiency in renal collecting ducts leads to PKD and promotes ciliary trafficking via increased KLC3.
BACKGROUND: Ciliogenesis-associated kinase 1 (CILK1) is a ciliary gene that localizes in primary cilia and regulates ciliary transport. Mutations in CILK1 cause various ciliopathies. However, the pathogenesis of CILK1-deficient kidney disease is unknown. METHODS: To examine whether CILK1 deficiency causes PKD accompanied by abnormal cilia, we generated mice with deletion of Cilk1 in cells of the renal collecting duct. A yeast two-hybrid system and coimmunoprecipitation (co-IP) were used to identify a novel regulator, kinesin light chain-3 (KLC3), of ciliary trafficking and cyst progression in the Cilk1-deficient model. Immunocytochemistry and co-IP were used to examine the effect of KLC3 on ciliary trafficking of the IFT-B complex and EGFR. We evaluated the effects of these genes on ciliary trafficking and cyst progression by modulating CILK1 and KLC3 expression levels. RESULTS: CILK1 deficiency leads to PKD accompanied by abnormal ciliary trafficking. KLC3 interacts with CILK1 at cilia bases and is increased in cyst-lining cells of CILK1-deficient mice. KLC3 overexpression promotes ciliary recruitment of IFT-B and EGFR in the CILK1 deficiency condition, which contributes to the ciliary defect in cystogenesis. Reduction in KLC3 rescued the ciliary defects and inhibited cyst progression caused by CILK1 deficiency. CONCLUSIONS: Our findings suggest that CILK1 deficiency in renal collecting ducts leads to PKD and promotes ciliary trafficking via increased KLC3.
Authors: Yan Y Yip; Stefano Pernigo; Anneri Sanger; Mengjia Xu; Maddy Parsons; Roberto A Steiner; Mark P Dodding Journal: Proc Natl Acad Sci U S A Date: 2016-02-16 Impact factor: 11.205
Authors: Yu Mi Woo; Do Yeon Kim; Nam Jin Koo; Yong-Min Kim; Sunyoung Lee; Je Yeong Ko; Yubin Shin; Bo Hye Kim; Hyowon Mun; Seonju Choi; Eun Ji Lee; Jeong-Oh Shin; Eun Young Park; Jinwoong Bok; Jong Hoon Park Journal: Sci Rep Date: 2017-10-26 Impact factor: 4.379