Cargando…

Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis

BACKGROUND: Morphogenetic events that shape the Drosophila melanogaster 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 s...

Descripción completa

Detalles Bibliográficos
Autores principales: Zúñiga, Alejandro, Hödar, Christian, Hanna, Patricia, Ibáñez, Freddy, Moreno, Pablo, Pulgar, Rodrigo, Pastenes, Luis, González, Mauricio, Cambiazo, Verónica
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2761875/
https://www.ncbi.nlm.nih.gov/pubmed/19772636
http://dx.doi.org/10.1186/1741-7007-7-61
_version_ 1782172864824213504
author Zúñiga, Alejandro
Hödar, Christian
Hanna, Patricia
Ibáñez, Freddy
Moreno, Pablo
Pulgar, Rodrigo
Pastenes, Luis
González, Mauricio
Cambiazo, Verónica
author_facet Zúñiga, Alejandro
Hödar, Christian
Hanna, Patricia
Ibáñez, Freddy
Moreno, Pablo
Pulgar, Rodrigo
Pastenes, Luis
González, Mauricio
Cambiazo, Verónica
author_sort Zúñiga, Alejandro
collection PubMed
description BACKGROUND: Morphogenetic events that shape the Drosophila melanogaster 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 D. melanogaster embryogenesis. We studied the spatiotemporal activity of these genes in five different intervals covering 12 stages of embryogenesis. RESULTS: 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. In situ 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 Drosophila 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. CONCLUSION: Our data have yielded a list of developmentally regulated D. melanogaster 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 D. melanogaster embryogenesis. These genes are also good candidates for additional targeted functional analyses similar to those we conducted for CG6234. See related minireview by Vichas and Zallen:
format Text
id pubmed-2761875
institution National Center for Biotechnology Information
language English
publishDate 2009
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-27618752009-10-15 Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis Zúñiga, Alejandro Hödar, Christian Hanna, Patricia Ibáñez, Freddy Moreno, Pablo Pulgar, Rodrigo Pastenes, Luis González, Mauricio Cambiazo, Verónica BMC Biol Research Article BACKGROUND: Morphogenetic events that shape the Drosophila melanogaster 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 D. melanogaster embryogenesis. We studied the spatiotemporal activity of these genes in five different intervals covering 12 stages of embryogenesis. RESULTS: 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. In situ 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 Drosophila 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. CONCLUSION: Our data have yielded a list of developmentally regulated D. melanogaster 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 D. melanogaster embryogenesis. These genes are also good candidates for additional targeted functional analyses similar to those we conducted for CG6234. See related minireview by Vichas and Zallen: BioMed Central 2009-09-22 /pmc/articles/PMC2761875/ /pubmed/19772636 http://dx.doi.org/10.1186/1741-7007-7-61 Text en Copyright © 2009 Zúñiga et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Zúñiga, Alejandro
Hödar, Christian
Hanna, Patricia
Ibáñez, Freddy
Moreno, Pablo
Pulgar, Rodrigo
Pastenes, Luis
González, Mauricio
Cambiazo, Verónica
Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis
title Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis
title_full Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis
title_fullStr Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis
title_full_unstemmed Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis
title_short Genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during Drosophila melanogaster embryogenesis
title_sort genes encoding novel secreted and transmembrane proteins are temporally and spatially regulated during drosophila melanogaster embryogenesis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2761875/
https://www.ncbi.nlm.nih.gov/pubmed/19772636
http://dx.doi.org/10.1186/1741-7007-7-61
work_keys_str_mv AT zunigaalejandro genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT hodarchristian genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT hannapatricia genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT ibanezfreddy genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT morenopablo genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT pulgarrodrigo genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT pastenesluis genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT gonzalezmauricio genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis
AT cambiazoveronica genesencodingnovelsecretedandtransmembraneproteinsaretemporallyandspatiallyregulatedduringdrosophilamelanogasterembryogenesis