Intracellular attenuation of BMP signaling via CKIP-1/Smurf1 is essential during neural crest induction
Abstract
The neural crest is induced at the neural plate border during gastrulation by combined bone morphogenetic protein (BMP), fibroblast growth factor (FGF), and Wnt signaling. While intermediate BMP levels are critical for this induction, secreted BMP inhibitors are largely absent from the neural plate border. Here, we propose a morphogen model in which intracellular attenuation of BMP signaling sets the required intermediate levels to maintain neural crest induction. We show that the scaffold protein casein kinase interacting protein 1 (CKIP-1) and ubiquitin ligase Smad ubiquitin regulatory factor 1 (Smurf1) are coexpressed with BMP4 at the chick neural plate border. Knockdown of CKIP-1 during a critical period between gastrulation and neurulation causes neural crest loss. Consistent with specific BMP modulation, CKIP-1 loss suppresses phospho-Smads 1/5/8 (pSmad1/5/8) and BMP reporter output but has no effect on Wnt signaling; Smurf1 overexpression (OE) acts similarly. Epistasis experiments further show that CKIP-1 rescues Smurf1-mediated neural crest loss. The results support a model in which CKIP-1 suppresses Smurf1-mediated degradation of Smads, uncovering an intracellular mechanism for attenuation of BMP signaling to the intermediate levels required for maintenance of neural crest induction.
Additional Information
© 2018 Piacentino, Bronner. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: October 4, 2017; Accepted: June 13, 2018; Published: June 27, 2018. Data Availability Statement: All relevant experimental data are within the paper, and underlying data can be found in S1 Data. Chicken CKIP-1 (Accession #KY982274) and Smurf1 (Accession #KY982275) cDNA sequences have been uploaded to Genbank. Funding: Eunice Kennedy Shriver National Institute of Child Health and Human Development https://www.nih.gov/ (grant number F32 HD088022). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Eunice Kennedy Shriver National Institute of Child Health and Human Development https://www.nih.gov/ (grant number P01 HD037105). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. National Institute of Dental and Craniofacial Research https://www.nih.gov/ (grant number R01 DE024157). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We would like to acknowledge Elisa Martí, Anna-Katerina Hadjantonakis, and Gerald Thomsen for sharing reagents. We also thank Steven Wilbert, Johanna Tan-Cabugao, and Harry Choi for technical assistance and Erica Hutchins, Stephen Green, and Megan Martik for valuable discussion. Confocal imaging was performed in the Biological Imaging Facility with the support of the Caltech Beckman Institute and the Arnold and Mabel Beckman Foundation. Author Contributions: Conceptualization: Michael L. Piacentino, Marianne E. Bronner. Data curation: Michael L. Piacentino. Formal analysis: Michael L. Piacentino. Funding acquisition: Michael L. Piacentino, Marianne E. Bronner. Investigation: Michael L. Piacentino. Methodology: Michael L. Piacentino. Resources: Michael L. Piacentino. Supervision: Marianne E. Bronner. Visualization: Michael L. Piacentino. Writing ± original draft: Michael L. Piacentino, Marianne E. Bronner. Writing ± review & editing: Michael L. Piacentino, Marianne E. Bronner. The authors have declared that no competing interests exist.Attached Files
Published - journal.pbio.2004425.pdf
Supplemental Material - journal.pbio.2004425.s001.tif
Supplemental Material - journal.pbio.2004425.s002.tif
Supplemental Material - journal.pbio.2004425.s003.tif
Supplemental Material - journal.pbio.2004425.s004.tif
Supplemental Material - journal.pbio.2004425.s005.tif
Supplemental Material - journal.pbio.2004425.s006.xlsx
Supplemental Material - journal.pbio.2004425.s007.xlsx
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Additional details
- PMCID
- PMC6039030
- Eprint ID
- 87504
- Resolver ID
- CaltechAUTHORS:20180702-081840117
- F32 HD088022
- NIH Postdoctoral Fellowship
- P01 HD037105
- NIH
- R01 DE024157
- NIH
- Caltech Beckman Institute
- Arnold and Mabel Beckman Foundation
- Created
-
2018-07-03Created from EPrint's datestamp field
- Updated
-
2023-06-01Created from EPrint's last_modified field