title: Developmental control of nuclear morphogenesis and anchoring by charleston, identified in a functional genomic screen of Drosophila cellularisation.
abstract: Morphogenesis of epithelial tissues relies on the precise developmental control of cell polarity and architecture. In the early Drosophila embryo, the primary epithelium forms during cellularisation, following a tightly controlled genetic programme where specific sets of genes are upregulated. Some of them, for example, control membrane invagination between the nuclei anchored at the apical surface of the syncytium. We used microarrays to describe the global programme of gene expression underlying cellularisation and identified distinct classes of upregulated genes during this process. Fifty-seven genes were then tested functionally by RNAi. We found six genes affecting various aspects of cellular architecture: membrane growth, organelle transport or organisation and junction assembly. We focus here on charleston (char), a new regulator of nuclear morphogenesis and of apical nuclear anchoring. In char-depleted embryos, the nuclei fail to maintain their elongated shape and, instead, become rounded. In addition, together with a disruption of the centrosome-nuclear envelope interaction, the nuclei lose their regular apical anchoring. These nuclear defects perturb the regular columnar organisation of epithelial cells in the embryo. Although microtubules are required for both nuclear morphogenesis and anchoring, char does not control microtubule organisation and association to the nuclear envelope. We show that Char is lipid anchored at the nuclear envelope by a farnesylation group, and localises at the inner nuclear membrane together with Lamin. Our data suggest that Char forms a scaffold that regulates nuclear architecture to constrain nuclei in tight columnar epithelial cells. The upregulation of Char during cellularisation and gastrulation reveals the existence of an as yet unknown developmental control of nuclear morphology and anchoring in embryonic epithelia.
authors.affiliation: Institut de Biologie du Développement de Marseille (IBDM de Génétique et de Physiologie du Développement (LGPD), UMR6545 CNRS-Université de la Méditerrannée. Campus de Luminy case 907, Marseille 13288 cedex9, France.
authors.forename: Fanny
authors.initials: F
authors.lastname: Pilot
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Philippe
authors.forename: Céline
authors.initials: C
authors.lastname: Lemmers
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Chauvin
authors.forename: Thomas
authors.initials: T
authors.lastname: Lecuit
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000037:antagonists & inhibitors / Q000378:metabolism; D000818:Animals; D002467:Cell Nucleus / Q000502:physiology* / Q000648:ultrastructure; D002478:Cells, Cultured; D018385:Centrosome / Q000502:physiology / Q000648:ultrastructure; D029721:Drosophila Proteins / Q000235:genetics / Q000502:physiology*; D004331:Drosophila melanogaster / Q000196:embryology / Q000502:physiology*; D004625:Embryo, Nonmammalian / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D007365:Intercellular Junctions / Q000502:physiology / Q000648:ultrastructure; D034882:Lamins / Q000502:physiology; D008715:Methionine / Q000031:analogs & derivatives / Q000494:pharmacology; D008870:Microtubules / Q000502:physiology / Q000648:ultrastructure; D009024:Morphogenesis; D009685:Nuclear Envelope / Q000502:physiology / Q000648:ultrastructure; D020411:Oligonucleotide Array Sequence Analysis
chemical_list: D029721:Drosophila Proteins; C096856:FTI 277; D034882:Lamins; D008715:Methionine; D019883:Alkyl and Aryl Transferases