Four distinct regulatory regions of the cut locus and their effect on cell type specification in Drosophila. It is possible that Heph may affect the activity of the gypsy insulator, since overexpression of Heph did not produce an appreciable alteration of the ct 53d wing phenotype. Note that Cut is still expressed in some sensory precursor cells outside of the CGal4 expression domain asterisks. Lola regulates midline crossing of CNS axons in Drosophila. In contrast to the ct 53d allele interaction, disruptions in BRM complex activity suppressed the discontinuities in the wing bristles of the gypsy ct K allele.
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Consistent with the ability to interact with cut during wing development, lola mRNA is expressed ubiquitously in the imaginal wing disc. During embryonic and pupal development Cut is specifically expressed in the neuroepithelial-derived sensory organ precursor cells SOP cells from which emerge the lineage-related cells of individual ES organs B lochlinger et al. G ptc-Gal4 drives expression along the anteroposterior axis of the wing disc.
HA is a non-enhancer of phenotype of ct 6. A In ct 53d mutants, Cut expression is reduced at the presumptive wing tip open arrowhead. Parameters of in vivo RNAi using Valium.
As part of the deficiency screen, two deficiencies, Df 3R red1 and Df 3R e-N19respectively covering Su Hw and mod mdg4were tested for an interaction with ct K.
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In homozygous lola mutant clones located adjacent to or bisecting the wing margin, neither Cut expression nor the morphology 9c6 wing-margin bristles is disrupted data not shown. Characterization of a novel discless mutant in D. UAS has visible phenotype, enhanceable by lola EP The GS-V stock was a generous gift from T.
Standard error of mean is given. This is in contrast to the observed c6 of Wg into cells adjacent to the boundary, indicating that the induction of Wg is independent of Cut.
Degeneration of margin cells prefigures the development of several rows of ES organs arrayed along the anterior wing margin. Distinct expression patterns detected within individual tissues by the GAL4 enhancer trap technique. Similarly, the chromosome deficiencies Df 2R vg -B, Df 2R Px4and Df 2R nap1covering loci previously x96 to encode positive regulators of Cut expression tal4 including the genes vestigialChipand Nipped-Brespectively, were tested for an interaction with ct K.
Hairpin constructs may produce a higher abundance of dsRNAs in males because they develop slightly more slowly than females. Further, to evaluate the differences in potency between different hairpins directed against the ggal4 genes, we generated two hairpins per gene when possible.
Identification of Genetic Loci That Interact With cut During Drosophila Wing-Margin Development
Overexpression of eIF-4E and eIF-4A may relieve putative cell growth or survival deficits c996 with the loss of cut activity by enhancing the translation of Cut target genes. HA is an enhancer of visible phenotype of ct 53d.
To determine a Wg-independent requirement for Cut in wing-margin development, we prevented degeneration of margin tissue in cut mutants by 1 maintaining Wg expression ectopically and 2 preventing apoptotic cell death through the misexpression of the baculovirus caspase inhibitor p Brand AH, Perrimon N.
UAS has wing phenotype, non-enhanceable by pan 13a. The regulation of the distal cut wing enhancer requires the activity of both enhancer-binding and enhancer-facilitator proteins.
Mathey-Prevot for critical input on the project, B. The differential interaction of group B candidates with various cut alleles likely reflects either direct or indirect effects on both gypsy insulator activity and cut wing-enhancer-mediated transcription.
Vector and parameters for targeted transgenic RNA interference in Drosophila melanogaster
Finally, the high rate of false negatives, resulting from random integration into poorly expressed loci, is automatically eliminated by inserting RNAi constructs into an optimal site such as attP2.
This could account for why we did not identify Df 3R e-N19 as a dominant suppressor of ct K. The main advantage of the method, in addition cc96 its relatively simple design and fast execution time, is that it allows spatial and temporal control of the knockdown construct, which is essential for characterizing genes with pleiotropic functions. In the developing wing, cut is required for proper patterning of the wing margin via complex interactions with multiple signaling pathways, including the Wingless Wg and Notch pathways M icchelli et al.
The genetic interaction data with ct 53d are summarized in Table 2. C996 data do not distinguish between these possibilities. Genetic analysis of brahma: UAS has wing margin bristle phenotype, enhanceable by N nd E Cut expression is reduced at val4 presumptive distal wing tip open arrowhead in ct 53d mutants.