title: Xenopus laevis nucleotide binding protein 1 (xNubp1) is important for convergent extension movements and controls ciliogenesis via regulation of the actin cytoskeleton.
abstract: Nucleotide binding protein 1 (Nubp1) is a highly conserved phosphate loop (P-loop) ATPase involved in diverse processes including iron-sulfur protein assembly, centrosome duplication and lung development. Here, we report the cloning, expression and functional characterization of Xenopus laevis Nubp1. We show that xNubp1 is expressed maternally, displays elevated expression in neural tissues and is required for convergent extension movements and neural tube closure. In addition, xNubp1knockdown leads to defective ciliogenesis of the multi-ciliated cells of the epidermis as well as the monociliated cells of the gastrocoel roof plate. Specifically, xNubp1 is required for basal body migration, spacing and docking in multi-ciliated cells and basal body positioning and axoneme elongation in monociliated gastrocoel roof plate cells. Live imaging of the different pools of actin and basal body migration during the process of ciliated cell intercalation revealed that two independent pools of actin are present from the onset of cell intercalation; an internal network surrounding the basal bodies, anchoring them to the cell cortex and an apical pool of punctate actin which eventually matures into the characteristic apical actin network. We show that xNubp1 colocalizes with the apical actin network of multiciliated cells and that problems in basal body transport in xNubp1 morphants are associated with defects of the internal network of actin, while spacing and polarity issues are due to a failure of the apical and sub-apical actin pools to mature into a network. Effects of xNubp1 knockdown on the actin cytoskeleton are independent of RhoA localization and activation, suggesting that xNubp1 may have a direct role in the regulation of the actin cytoskeleton.
authors.affiliation: Department of Biological Sciences, University of Cyprus, Nicosia 2109, Cyprus. Ioannou.andriani@ucy.ac.cy
authors.forename: Andriani
authors.initials: A
authors.lastname: Ioannou
authors.forename: Niovi
authors.initials: N
authors.lastname: Santama
authors.forename: Paris A
authors.initials: PA
authors.lastname: Skourides
mesh_terms: D008841:Actin Cytoskeleton / Q000502:physiology*; D000818:Animals; D002465:Cell Movement; D002923:Cilia / Q000502:physiology*; D005260:Female; D019204:GTP-Binding Proteins / Q000502:physiology*; D009024:Morphogenesis; D029867:Xenopus Proteins / Q000502:physiology*; D014982:Xenopus laevis / Q000196:embryology*
chemical_list: C583340:Nubp1 protein, Xenopus; D029867:Xenopus Proteins; D019204:GTP-Binding Proteins
keywords: Actin; Basal bodies; Ciliogenesis; Convergent extension; Nubp1; Xenopus