Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesis
<p>Abstract</p> <p>Background</p> <p>Morphogenetic events that shape the <it>Drosophila melanogaster </it>embryo are tightly controlled by a genetic program in which specific sets of genes are up-regulated. We used a suppressive subtractive hybridization pro...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2009-09-01
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| Series: | BMC Biology |
| Online Access: | http://www.biomedcentral.com/1741-7007/7/61 |
| _version_ | 1811251977607184384 |
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| author | González Mauricio Pastenes Luis Pulgar Rodrigo Moreno Pablo Ibáñez Freddy Hanna Patricia Hödar Christian Zúñiga Alejandro Cambiazo Verónica |
| author_facet | González Mauricio Pastenes Luis Pulgar Rodrigo Moreno Pablo Ibáñez Freddy Hanna Patricia Hödar Christian Zúñiga Alejandro Cambiazo Verónica |
| author_sort | González Mauricio |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Morphogenetic events that shape the <it>Drosophila melanogaster </it>embryo are tightly controlled by a genetic program in which specific sets of genes are up-regulated. We used a suppressive subtractive hybridization procedure to identify a group of developmentally regulated genes during early stages of <it>D. melanogaster </it>embryogenesis. We studied the spatiotemporal activity of these genes in five different intervals covering 12 stages of embryogenesis.</p> <p>Results</p> <p>Microarrays were constructed to confirm induction of expression and to determine the temporal profile of isolated subtracted cDNAs during embryo development. We identified a set of 118 genes whose expression levels increased significantly in at least one developmental interval compared with a reference interval. Of these genes, 53% had a phenotype and/or molecular function reported in the literature, whereas 47% were essentially uncharacterized. Clustering analysis revealed demarcated transcript groups with maximum gene activity at distinct developmental intervals. <it>In situ </it>hybridization assays were carried out on 23 uncharacterized genes, 15 of which proved to have spatiotemporally restricted expression patterns. Among these 15 uncharacterized genes, 13 were found to encode putative secreted and transmembrane proteins. For three of them we validated our protein sequence predictions by expressing their cDNAs in <it>Drosophila </it>S2R+ cells and analyzed the subcellular distribution of recombinant proteins. We then focused on the functional characterization of the gene CG6234. Inhibition of CG6234 by RNA interference resulted in morphological defects in embryos, suggesting the involvement of this gene in germ band retraction.</p> <p>Conclusion</p> <p>Our data have yielded a list of developmentally regulated <it>D. melanogaster </it>genes and their expression profiles during embryogenesis and provide new information on the spatiotemporal expression patterns of several uncharacterized genes. In particular, we recovered a substantial number of unknown genes encoding putative secreted and transmembrane proteins, suggesting new components of signaling pathways that might be incorporated within the existing regulatory networks controlling <it>D. melanogaster </it>embryogenesis. These genes are also good candidates for additional targeted functional analyses similar to those we conducted for CG6234.</p> <p>See related minireview by Vichas and Zallen: <url>http://www.jbiol.com/content/8/8/76</url></p> |
| first_indexed | 2024-04-12T16:28:03Z |
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| id | doaj.art-1b1ba19fd8754fc6b6637bce46e547dc |
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| issn | 1741-7007 |
| language | English |
| last_indexed | 2024-04-12T16:28:03Z |
| publishDate | 2009-09-01 |
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| spelling | doaj.art-1b1ba19fd8754fc6b6637bce46e547dc2022-12-22T03:25:16ZengBMCBMC Biology1741-70072009-09-01716110.1186/1741-7007-7-61Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesisGonzález MauricioPastenes LuisPulgar RodrigoMoreno PabloIbáñez FreddyHanna PatriciaHödar ChristianZúñiga AlejandroCambiazo Verónica<p>Abstract</p> <p>Background</p> <p>Morphogenetic events that shape the <it>Drosophila melanogaster </it>embryo are tightly controlled by a genetic program in which specific sets of genes are up-regulated. We used a suppressive subtractive hybridization procedure to identify a group of developmentally regulated genes during early stages of <it>D. melanogaster </it>embryogenesis. We studied the spatiotemporal activity of these genes in five different intervals covering 12 stages of embryogenesis.</p> <p>Results</p> <p>Microarrays were constructed to confirm induction of expression and to determine the temporal profile of isolated subtracted cDNAs during embryo development. We identified a set of 118 genes whose expression levels increased significantly in at least one developmental interval compared with a reference interval. Of these genes, 53% had a phenotype and/or molecular function reported in the literature, whereas 47% were essentially uncharacterized. Clustering analysis revealed demarcated transcript groups with maximum gene activity at distinct developmental intervals. <it>In situ </it>hybridization assays were carried out on 23 uncharacterized genes, 15 of which proved to have spatiotemporally restricted expression patterns. Among these 15 uncharacterized genes, 13 were found to encode putative secreted and transmembrane proteins. For three of them we validated our protein sequence predictions by expressing their cDNAs in <it>Drosophila </it>S2R+ cells and analyzed the subcellular distribution of recombinant proteins. We then focused on the functional characterization of the gene CG6234. Inhibition of CG6234 by RNA interference resulted in morphological defects in embryos, suggesting the involvement of this gene in germ band retraction.</p> <p>Conclusion</p> <p>Our data have yielded a list of developmentally regulated <it>D. melanogaster </it>genes and their expression profiles during embryogenesis and provide new information on the spatiotemporal expression patterns of several uncharacterized genes. In particular, we recovered a substantial number of unknown genes encoding putative secreted and transmembrane proteins, suggesting new components of signaling pathways that might be incorporated within the existing regulatory networks controlling <it>D. melanogaster </it>embryogenesis. These genes are also good candidates for additional targeted functional analyses similar to those we conducted for CG6234.</p> <p>See related minireview by Vichas and Zallen: <url>http://www.jbiol.com/content/8/8/76</url></p>http://www.biomedcentral.com/1741-7007/7/61 |
| spellingShingle | González Mauricio Pastenes Luis Pulgar Rodrigo Moreno Pablo Ibáñez Freddy Hanna Patricia Hödar Christian Zúñiga Alejandro Cambiazo Verónica Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesis BMC Biology |
| title | Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesis |
| title_full | Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesis |
| title_fullStr | Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesis |
| title_full_unstemmed | Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesis |
| title_short | Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during <it>Drosophila melanogaster </it>embryogenesis |
| title_sort | genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during it drosophila melanogaster it embryogenesis |
| url | http://www.biomedcentral.com/1741-7007/7/61 |
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