Signalling Pathways in Development and Human Disease: A Drosophila Wing Perspective

Molnar, Cristina; Resnik-Docampo, Martin; Organista, María F.; Martı́n, Mercedes; Hevia, Covadonga F.; Celis, José F. de

doi:10.5772/23858

Cited by 10 publications

(15 citation statements)

References 253 publications

(173 reference statements)

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“…In 4 of these cases there was only one candidate gene, and the failure of the corresponding RNAi to affect the overexpression phenotype under the conditions in which we used them suggests that these RNAi were not effective enough to reduce the levels of the overexpressed transcript. The fraction of false negatives is similar to that found in large-scale screens using the UASRNAi collection (Mummery-Widmer et al 2009). In addition, we also found 8 cases in which two different RNAi suppress the overexpression phenotype of a particular P-GS insertion (Table 1, see examples in Figure 2, H-I999).…”

supporting

confidence: 56%

“…More likely, the false-negative cases of nonrescue were due to inefficient gene silencing caused by either insufficient levels of RNAi expression driven by the UAS-RNAi constructs or by ineffective RNA interference. This result is not surprising, because an estimated fraction of 29% of UAS-RNAi lines is inefficient to cause gene silencing (Mummery-Widmer et al 2009). In addition, we also found a fraction of cases (9%) in which two candidate genes rescued to some extent the Gal4/P-GS phenotype, which we assume corresponds to cases in which two neighbor genes need to be overexpressed simultaneously to result in an overexpression phenotype.…”

Section: Discussionmentioning

confidence: 95%

“…For some genes, such as those involved in signaling, or for transcription factors affecting cell-fate choices, there is a good complementarity between the loss-and gain-of expression phenotype (Molnar et al 2011). In this manner, mutations reducing EGFR activity, for example, cause loss of veins, whereas the increase in EGFR causes the formation of ectopic veins (Sturtevant and Bier 1995;Guichard et al 1999).…”

Section: Loss-of-function Phenotype Of Candidate Genesmentioning

confidence: 99%

“…For these reasons, gain-of-function screens constitute an effective entry point from which to characterize the requirements and molecular function of a gene in a particular developmental process. In addition, and for those genes affecting cell-fate choices such as several transcription factors and signaling molecules, there is a reasonably good complementarity between the loss-of-function and the gain-of-function phenotypes (Molnar et al 2011). This complementarity allows refininement of the function assigned to the gene, as well as identification of additional components of signaling pathways by the consequences of their overexpression.…”

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confidence: 99%

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Genetic Annotation of Gain-Of-Function Screens Using RNA Interference and in Situ Hybridization of Candidate Genes in the Drosophila Wing

et al. 2012

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Gain-of-function screens in Drosophila are an effective method with which to identify genes that affect the development of particular structures or cell types. It has been found that a fraction of 2-10% of the genes tested, depending on the particularities of the screen, results in a discernible phenotype when overexpressed. However, it is not clear to what extent a gain-of-function phenotype generated by overexpression is informative about the normal function of the gene. Thus, very few reports attempt to correlate the loss-and overexpression phenotype for collections of genes identified in gain-of-function screens. In this work we use RNA interference and in situ hybridization to annotate a collection of 123 P-GS insertions that in combination with different Gal4 drivers affect the size and/or patterning of the wing. We identify the gene causing the overexpression phenotype by expressing, in a background of overexpression, RNA interference for the genes affected by each P-GS insertion. Then, we compare the loss and gainof-function phenotypes obtained for each gene and relate them to its expression pattern in the wing disc. We find that 52% of genes identified by their overexpression phenotype are required during normal development. However, only in 9% of the cases analyzed was there some complementarity between the gain-and loss-of-function phenotype, suggesting that, in general, the overexpression phenotypes would not be indicative of the normal requirements of the gene.

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supporting

confidence: 56%

Section: Discussionmentioning

confidence: 95%

Section: Loss-of-function Phenotype Of Candidate Genesmentioning

confidence: 99%

mentioning

confidence: 99%

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Genetic Annotation of Gain-Of-Function Screens Using RNA Interference and in Situ Hybridization of Candidate Genes in the Drosophila Wing

et al. 2012

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“…Although this reported on the adverse consequences of miRNA misexpression, it did not provide substantial phenotypic information. We therefore turned to the adult wing, as wing development requires the coordinate function of multiple signaling pathways (Molnar et al, 2011), and even minor alterations in wing development are visible under the dissecting microscope.…”

Section: Research Articlementioning

confidence: 99%

A genome-wide transgenic resource for conditional expression ofDrosophilamicroRNAs

et al. 2012

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SUMMARYmicroRNAs (miRNAs) are endogenous short RNAs that mediate vast networks of post-transcriptional gene regulation. Although computational searches and experimental profiling provide evidence for hundreds of functional targets for individual miRNAs, such data rarely provide clear insight into the phenotypic consequences of manipulating miRNAs in vivo. We describe a genome-wide collection of 165 Drosophila miRNA transgenes and find that a majority induced specific developmental defects, including phenocopies of mutants in myriad cell-signaling and patterning genes. Such connections allowed us to validate several likely targets for miRNA-induced phenotypes. Importantly, few of these phenotypes could be predicted from computationally predicted target lists, thus highlighting the value of whole-animal readouts of miRNA activities. Finally, we provide an example of the relevance of these data to miRNA loss-of-function conditions. Whereas misexpression of several K box miRNAs inhibited Notch pathway activity, reciprocal genetic interaction tests with miRNA sponges demonstrated endogenous roles of the K box miRNA family in restricting Notch signaling. In summary, we provide extensive evidence that misexpression of individual miRNAs often induces specific mutant phenotypes that can guide their functional study. By extension, these data suggest that the deregulation of individual miRNAs in other animals may frequently yield relatively specific phenotypes during disease conditions.

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Functional requirements of protein kinases and phosphatases in the development of the Drosophila melanogaster wing

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Esteban

López-Varea

et al. 2021

G3 Genes|Genomes|Genetics

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Protein kinases and phosphatases constitute a large family of conserved enzymes that control a variety of biological processes by regulating the phosphorylation state of target proteins. They play fundamental regulatory roles during cell cycle progression and signaling, among other key aspects of multicellular development. The complement of protein kinases and phosphatases includes approximately 326 members in Drosophila, and they have been the subject of several functional screens searching for novel components of signaling pathways and regulators of cell division and survival. These approaches have been carried out mostly in cell cultures using RNA interference to evaluate the contribution of each protein in different functional assays, and have contributed significantly to assign specific roles to the corresponding genes. In this work we describe the results of an evaluation of the Drosophila complement of kinases and phosphatases using the wing as a system to identify their functional requirements in vivo. We also describe the results of several modifying screens aiming to identify among the set of protein kinases and phosphatases additional components or regulators of the activities of the Epidermal Growth Factor and Insulin receptors signaling pathways.

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Signalling Pathways in Development and Human Disease: A Drosophila Wing Perspective

Cited by 10 publications

References 253 publications

Genetic Annotation of Gain-Of-Function Screens Using RNA Interference and in Situ Hybridization of Candidate Genes in the Drosophila Wing

Genetic Annotation of Gain-Of-Function Screens Using RNA Interference and in Situ Hybridization of Candidate Genes in the Drosophila Wing

A genome-wide transgenic resource for conditional expression ofDrosophilamicroRNAs

Functional requirements of protein kinases and phosphatases in the development of the Drosophila melanogaster wing

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