RNA-binding protein Elavl1/HuR is required for maintenance of cranial neural crest specification

Abstract

Neural crest development is transcriptionally controlled via sequential activation of gene regulatory networks (GRNs). Recent evidence increasingly implicates a role for post-transcriptional regulation in modulating the output of these regulatory circuits. Using RNA-sequencing data from avian embryos to identify potential post-transcriptional regulators, we observed enrichment during early neural crest development of Elavl1, which encodes for the RNA-binding protein HuR. Immunohistochemical analyses revealed expression of HuR following establishment of the neural plate border. Perturbation of HuR resulted in premature neural crest delamination from the neural tube as well as significant reduction in transcripts associated with the neural crest specification GRN (Axud1 and FoxD3), phenotypes also observed with downregulation of the canonical Wnt inhibitor Draxin. RNA pulldown further shows that Draxin is a specific target of HuR. Importantly, overexpression of exogenous Draxin was able to rescue the cranial neural crest specification defects observed with HuR knockdown. Thus, HuR plays a critical a role in the maintenance of cranial neural crest specification, at least partially via Draxin mRNA stabilization. Together, these data highlight an important intersection of post-transcriptional regulation with modulation of the neural crest specification GRN.