Following the intracellular localization of the iab-8ncRNA of the bithorax complex using the MS2-MCP-GFP system.
Homeotic genes are aligned on the chromosome in the order of the segments that they specify along the antero-posterior axis of the fly. In general the genes affecting the more posterior segments repress the more anterior genes, a phenomenon known as "posterior dominance". There is however a noticeable exception to this rule in the central nervous system of Drosophila melanogaster where the posterior Abd-B gene does not repress the immediately more anterior abd-A gene. Instead, abd-A repression is accomplished by a 92 kb-long ncRNA (the iab-8ncRNA) that is transcribed from the large inter-genic region between abd-A and Abd-B. This iab-8ncRNA encodes a microRNA to repress abd-A and also a second redundant repression mechanism acting in cis and thought to be transcriptional interference with the abd-A promoter. Using in situ hybridization, a previous work suggested that the iab8ncRNA transcript forms discrete foci restricted to the nuclear periphery and that this localization may be important for its function. In order to better characterize the intra-cellular localization of the iab-8ncRNA we used the MS2-MCP system, which allows fluorescent labeling of RNA in cells and relies on the interaction between GFP-tagged MS2 coat protein (MCP-GFP) and MS2 RNA stem loops. Our results indicate that the large foci seen in previous studies correspond to the site of iab8ncRNA transcription and that the foci seen may simply be an indication of the level of transcription at the locus. We find no evidence to suggest that this localization is important for its function on abd-A repression. We discuss the idea that the iab-8ncRNA may be a relic of a more general ancient mechanism of posterior dominance during the emergence of the hox clusters that was mediated by transcriptional interference.
Department of Genetic and Evolution, University of Geneva, 30 Quai E. Ansermet, 1211 Geneva-4, Switzerland.
Ghislaine
G
Arib
Department of Genetic and Evolution, University of Geneva, 30 Quai E. Ansermet, 1211 Geneva-4, Switzerland.
Fabienne
F
Cléard
Department of Genetic and Evolution, University of Geneva, 30 Quai E. Ansermet, 1211 Geneva-4, Switzerland.
Robert K
RK
Maeda
Department of Genetic and Evolution, University of Geneva, 30 Quai E. Ansermet, 1211 Geneva-4, Switzerland. Electronic address: Francois.Karch@unige.ch.
François
F
Karch
D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D005801:Genes, Homeobox / Q000235:genetics; D017344:Genes, Insect / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
D029721:Drosophila Proteins; D035683:MicroRNAs; C580822:iab-4 microRNA, Drosophila; D049452:Green Fluorescent Proteins
Bithorax; Hox genes; MicroRNA; Posterior dominance; ncRNA