Figure 1.
(A, B) Images of wild type (w1118) straight wing and SIN3 KD (Ser-GAL4 X UAS-SIN3RNAi) curved wing Drosophila as indicated. (C) SIN3 KD/CyO-Ras females were crossed to males heterozygous for either a deletion that removed multiple genes (phase I and II) or either a loss of function allele or RNAi line of a single gene (phase III) balanced over a third chromosome balancer that was TM3-Sb, TM3-Ser, TM2-Ubx or TM6-Tb. The resulting progeny that are Sin3A knockdown and carry a third chromosome deletion were scored for the curved wing phenotype. In phase I of the screen, each deletion on the third chromosome was associated with a gene resulting in yellow to orange eye color. In the case of phase II, all the deletions tested were in a w background. The majority of RNAi lines were homozygous and the cross yielded progeny of two genotypes.
Table 1.
Genes involved in multiple cellular processes genetically interact with Sin3A.
Figure 2.
Some genes along the third chromosome are required for normal wing development and interact with Sin3A.
Images of representative wings from progeny of Ser-GAL4 X UAS-RNAi (left panels) and from SIN3 KD X UAS-RNAi (right panels) of the indicated gene.
Table 2.
Genes involved in negative regulation of the Wnt pathway genetically interact with Sin3A.
Figure 3.
Reduced expression of negative regulators of the Wnt signaling pathway results in abnormal wing morphology.
Images of representative wings from progeny of Ser-GAL4 X UAS-RNAi (left panels) and from SIN3 KD X UAS-RNAi (right panel) of the indicated gene. For tum and sgg, the wing phenotype of the double knockdown was the same as for the single gene knockdown. The phenotype of the ft, Sin3A double knockdown is similar to the SIN3 KD curved wing as shown.
Figure 4.
Loss of Sin3A results in down regulation of genes involved in the Wnt pathway.
qRT-PCR analysis of the mRNAs of the indicated genes. mRNA from control w1118 and Sin3A knockdown wing discs was reverse transcribed into cDNA to use as template in the PCR. Gene expression in Sin3A knockdown wing discs relative to w1118 is indicated. Expression was normalized to Taf1 and Pgk expression. n = 3. Error bars indicate standard deviation. (*) 0.006<p<0.02. Gray bars, Wnt targets. White bars, Wnt targets and effectors. Black bar, Wnt effector.
Table 3.
Sin3A interacts genetically with cell cycle regulators.
Figure 5.
Wing development is sensitive to reduced expression of cell cycle regulators.
Images of representative wings from progeny of Ser-GAL4 X UAS-RNAi of the indicated gene (left panels). For each of these genes, the wing phenotype of the double knockdown was the same as for the single gene knockdown except where noted (right panels). For cdc2c, images representing the variable phenotypes in the population are shown.
Table 4.
Components of the Mediator kinase module genetically interact with Sin3A.
Figure 6.
Two components of the Mediator accessory kinase module are important for wing morphology.
Images of representative wings from progeny of Ser-GAL4 X UAS-RNAi of the indicated gene. For CycC, the wing phenotype of the double knockdown was the same as for the single gene knockdown. The phenotype of the single and double knockdown phenotype with kto is shown.
Figure 7.
Multiple genes that reside along the third chromosome are required for normal wing morphology.
Images of wings from progeny of Ser-GAL4 X UAS-RNAi of the indicated gene. For each of these genes, the wing phenotype of the double knockdown was the same as for the single gene knockdown. In cases where the phenotype was variable in the population, multiple representative images are shown.