Ric-8A controls Cited2 subcellular localization and transcriptional programs during neural crest development

Neural crest (NC) cells are a highly migratory and multipotent population essential for vertebrate development, whose behavior depends on the integration of signaling pathways, transcriptional programs, and cytoskeletal dynamics. Here, we identify a previously unrecognized functional relationship between the signaling regulator Ric-8A and the transcriptional co-regulator Cited2 during NC development in Xenopus. Proteomic analyses and proximity ligation assays revealed Cited2 as a Ric-8A-associated factor, prompting us to investigate its role in NC biology. We show that xtcited2 is dynamically expressed during embryogenesis, with enrichment in neural and mesodermal derivatives, including cranial NC. At the cellular level, XtCited2 exhibits a dual nuclear-cytoplasmic distribution that is regulated by XtRic-8A. Gain- and loss-of-function experiments, together with rescue assays, demonstrate that XtRic-8A controls the subcellular localization of XtCited2 and influences XtCited2-dependent transcriptional programs. Single-cell transcriptomic analyses further revealed that co-expression of xtcited2 and xtric-8a defines a distinct transcriptional state enriched for genes associated with cytoskeletal organization, Rho GTPase signaling, cell migration, and cell-cycle regulation. Consistently, functional assays showed that XtRic-8A is required to maintain normal sox2 expression, linking this regulatory axis to transcriptional control of NC identity. Together, our findings uncover a regulatory mechanism by which XtRic-8A modulates XtCited2 function, connecting protein localization, transcriptional regulation, and NC cell behavior during vertebrate development.