NEDL2 regulates enteric nervous system and kidney development in its Nedd8 ligase activity-dependent manner.
The GDNF (Glial cell line-derived neurotrophic factor)/Ret/Akt signaling pathway is essential to the development of ENS (enteric nervous system) as well as kidney. We previously showed that the HECT-type E3 ligase NEDL2 (Nedd4-like ligase 2) is required for the ENS development by activating GDNF/Ret/Akt. However, the underlying mechanism remains unknown. Here we show that in addition to ENS, NEDL2 is also pivotal for kidney development since about 1/3 of Nedl2-deficient mice displayed postnatal unilateral or bilateral kidney hydronephrosis. Double knockout of Nedl1 and Nedl2 in mice leads to postnatal lethal within 2 weeks and the phenotypes resemble those of Nedl2 single knockout mice. Surprisingly, its close member NEDL1 is dispensable for ENS and kidney function and the reason is lack of NEDL1 expression in these systems during early development. Furthermore, biochemical analysis indicated that NEDL2 appears to act like a scaffold protein to recruit SHC, Grb2, PI3K (p110 and p85), PDK1 and Akt together to promote the signaling transduction. Intriguingly, we found that NEDL2 harbours intrinsic Nedd8 ligase activity with cysteine 1341 as the core site. NEDL2 upregulates GDNF-stimulated Akt activity dependent of its Nedd8 ligase activity but not its ubiquitin ligase activity. These findings demonstrate that NEDL2 but not NEDL1 is required for ENS and kidney development in a unique Nedd8 ligase-dependent manner.
School of Life Sciences, Tsinghua University, Beijing 100084, China.|State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.
Xiao
X
Qiu
School of Life Sciences, Tsinghua University, Beijing 100084, China.|State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.
Rongfei
R
Wei
Department of Experimental Pathology, Beijing Institute of Radiation Medicine, Beijing 100850, China.
Yang
Y
Li
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.|Institute of Cancer Stem Cell, Dalian Medical University, Dalian 116044, China.
Qiong
Q
Zhu
School of Life Sciences, Tsinghua University, Beijing 100084, China.
Cong
C
Xiong
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.
Yuhan
Y
Chen
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.
Yuan
Y
Zhang
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.
Kefeng
K
Lu
School of Life Sciences, Tsinghua University, Beijing 100084, China.|State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.
Fuchu
F
He
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Collaborative Innovation Center for Cancer Medicine, Beijing 100850, China.|Institute of Cancer Stem Cell, Dalian Medical University, Dalian 116044, China.
Lingqiang
L
Zhang
D000818:Animals; D017615:Enteric Nervous System / Q000254:growth & development / Q000378:metabolism*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007668:Kidney / Q000254:growth & development / Q000378:metabolism*; D061986:MCF-7 Cells; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D000075687:NEDD8 Protein / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051096:Proto-Oncogene Proteins c-ret / Q000378:metabolism; D034622:RNA Interference; D015398:Signal Transduction; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*
D051100:Glial Cell Line-Derived Neurotrophic Factor; D000075687:NEDD8 Protein; C513172:Nedd8 protein, mouse; C000602665:Hecw2 protein, mouse; D044767:Ubiquitin-Protein Ligases; D051096:Proto-Oncogene Proteins c-ret; D051057:Proto-Oncogene Proteins c-akt
GDNF/Ret/Akt pathway; Nedd8 ligase; kidney development; scaffold protein; ubiquitin ligase