Although a network of transcription factors that specifies neural crest identity in the ectoderm has been defined, expression of neural crest transcription factors does not guarantee eventual migration as a neural crest cell. While much work has gone into determining regulatory relationships within the transcription factor network, the ability of protein modifications like phosphorylation to modulate the function of neural crest regulatory factors and determine when and where they are active also has crucial implications. Paladin, which was previously classified as a phosphatase based on sequence similarity, is expressed in chick neural crest precursors and is maintained throughout their epithelial to mesenchymal transition and migration. Loss of Paladin delays the expression of transcription factors Snail2 and Sox10 in premigratory neural crest cells, but does not affect accumulation of FoxD3, Cad6B or RhoB, indicating that Paladin differentially modulates the expression of genes previously thought to be coregulated within the neural crest gene regulatory network. Both gain and loss of Paladin function result in disrupted neural crest migration, reinforcing the importance of precisely regulated phosphorylation for neural crest migration. Mutation of critical, catalytic cysteine residues within Paladin's predicted phosphatase active site motifs did not abolish the function of Paladin in the neural crest. Collectively, these data indicate that Paladin is an antiphosphatase that modulates the activity of specific neural crest regulatory factors during neural crest development. Our work identifies a novel regulator of phosphorylation status that provides an additional layer of regulation in the neural crest.
Bibliographical noteFunding Information:
We thank Carol Erickson, Paul Scotting, Andrea Streit, and Lisa Taneyhill for kind gifts of plasmids. Special thanks to Wuming Gong for assistance with statistical analyses. We are grateful to the members of the Gammill Lab and Yasuhiko Kawakami for their input over the course of this project. This work was supported by the National Institute of Dental and Craniofacial Research of the National Institutes of Health award number K22 DE15309 (to LSG) and Ruth L. Kirschstein National Research Service Award number F32 DE019973 (to JRA), and a Minnesota Medical Foundation Research Grant.
- Neural crest