Abstract
Emerging data indicate that actin dynamics is associated with ciliogenesis. However, the underlying mechanism remains unclear. Here we find that nuclear distribution gene C (NudC), an Hsp90 co-chaperone, is required for actin organization and dynamics. Depletion of NudC promotes cilia elongation and increases the percentage of ciliated cells. Further results show that NudC binds to and stabilizes cofilin 1, a key regulator of actin dynamics. Knockdown of cofilin 1 also facilitates ciliogenesis. Moreover, depletion of either NudC or cofilin 1 causes similar ciliary defects in zebrafish, including curved body, pericardial edema and defective left-right asymmetry. Ectopic expression of cofilin 1 significantly reverses the phenotypes induced by NudC depletion in both cultured cells and zebrafish. Thus, our data suggest that NudC regulates actin cytoskeleton and ciliogenesis by stabilizing cofilin 1.
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Acknowledgements
We thank Ping Wang for providing us the HA-ubiquitin expression vector; Xiao Huang for pCS107 vector. We are grateful to Yibin Kang, Lynn W Enquist and Lu Zhao for helpful suggestions; Lei Zhu, Yirong Zhang, Biao Zhu and Yijun Liu for technical assistance. We thank State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences and Chinese Academy of Sciences for instrument support. This work was supported by Ministry of Science and Technology of China (2012CB945004, 2013CB945603 and 2011CBA01001), National Natural Science Foundation of China (31125017, 31190063, 31100975, 31571446 and 31471259), Ministry of Education of China (20110101110103), Natural Scientific Foundation of Zhejiang Province, China (LY14C070001 and LY14C070002), the 111 Project (B13026), Department of Science and Technology of Zhejiang Province (2012C37089 and 2013TD13) and Zhejiang Provincial Program for the Cultivation of High-level Innovative Health talents.
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Supplementary information
Supplementary information, Figure S1
The cellular localization of NudC together with actin in RPE-1 cells. (PDF 249 kb)
Supplementary information, Figure S2
Ectopic expression of NudC rescues the actin defects induced by NudC depletion. (PDF 422 kb)
Supplementary information, Figure S3
Knockdown of NudC impairs collective cell migration. (PDF 1036 kb)
Supplementary information, Figure S4
Exogenous expression of NudC rescues the ciliary defects induced by NudC depletion. (PDF 252 kb)
Supplementary information, Figure S5
NudC suppresses ciliogenesis in HEK293T cells. (PDF 487 kb)
Supplementary information, Figure S6
NudC expression patterns during zebrafish development. (PDF 1143 kb)
Supplementary information, Figure S7
Zebrafish embryos injected with NudC MO-2 exhibit multiple ciliary defects. (PDF 1205 kb)
Supplementary information, Figure S8
Disruption of NudC by CRISPR/Cas9 causes ciliary defects. (PDF 561 kb)
Supplementary information, Figure S9
Cofilin 1 inhibits cilia assembly in HEK293T cells. (PDF 396 kb)
Supplementary information, Figure S10
Exogenous expression of cofilin 1 mRNA rescues the ciliary defects in cofilin 1 morphants. (PDF 978 kb)
Supplementary information, Figure S11
Cofilin 1 reverses the ciliary defects in zebrafish injected with NudC MO-2. (PDF 792 kb)
Supplementary information, Figure S12
Disruption of cofilin 1 by CRISPR/Cas9 induces ciliary phenotypes. (PDF 558 kb)
Supplementary information, Figure S13
Cofilin 1 partially rescues the enlarged pronephric duct in NudC morphants. (PDF 537 kb)
Supplementary information, Figure S14
The cellular localization of NudC in RPE-1 cells. (PDF 461 kb)
Supplementary information, Figure S15
Inhibition of Hsp90 activity suppresses ciliogenesis in mammalian cells. (PDF 315 kb)
Supplementary information, Figure S16
Inhibition of Hsp90 activity has no significant effect on the protein level of cofilin 1. (PDF 182 kb)
Supplementary information, Movie S1
Ciliary motility in pronephric duct of control morphant. Blue arrows show pronephric duct. (MPEG 4802 kb)
Supplementary information, Movie S2
Ciliary motility in pronephric duct of NudC morphant. Blue arrows indicate pronephric duct. (MPEG 4809 kb)
Supplementary information, Movie S3
Ciliary motility in pronephric duct of the embryo injected with NudC mRNA. Blue arrows show pronephric duct. (MPEG 4806 kb)
Supplementary information, Movie S4
Ciliary motility in pronephric duct of NudC morphant co-injected with NudC mRNA. Blue arrows show pronephric duct. (MPEG 4809 kb)
Supplementary information, Movie S5
Ciliary motility in pronephric duct of control morphant. Blue arrows show pronephric duct. (MPEG 4806 kb)
Supplementary information, Movie S6
Ciliary motility in pronephric duct of NudC morphant. Blue arrows indicate pronephric duct. (MPEG 4806 kb)
Supplementary information, Movie S7
Ciliary motility in pronephric duct of the embryo injected with cofilin 1 mRNA. Blue arrows show pronephric duct. (MPEG 4793 kb)
Supplementary information, Movie S8
Ciliary motility in pronephric duct of NudC morphant co-injected with cofilin 1 mRNA. Blue arrows show pronephric duct. (MPEG 4795 kb)
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Zhang, C., Zhang, W., Lu, Y. et al. NudC regulates actin dynamics and ciliogenesis by stabilizing cofilin 1. Cell Res 26, 239–253 (2016). https://doi.org/10.1038/cr.2015.152
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DOI: https://doi.org/10.1038/cr.2015.152
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