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Investigation of Dys Regulated Enhancers: Target Gene Activation and Molecular Mechanisms

This study explores the molecular mechanisms underlying the regulation of target genes by Dys and Tgo in a Drosophila model. We identify enhancer elements associated with CG13196 and CG15252, utilizing fusion cell gene profiling methods and the UAS-GFP reporter system. The findings indicate that Dys can activate specific enhancers at late developmental stages, with no evidence of transcriptional inhibition in early stages. Additionally, we demonstrate that Spalt is not necessary for CG15252 activation, suggesting alternative regulatory pathways may be involved.

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Investigation of Dys Regulated Enhancers: Target Gene Activation and Molecular Mechanisms

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  1. Identify dys fusion cell enhancer • Fusion cell gene profiling dys-gal4::UAS-CD8-GFP btl-gal4::UAS-CD8-GFP • Identify dys target gene dys-gal4::UAS-CD8-GFP dys-gal4::UAS-CD8-GFP ; dys/dys • Molecular mechanism of target gene regulation by Dys::Tgo • Dys::Tgo activation of CG13196 • Dys:Tgo activation of CG15252 • Identify mbo and CG15252 enhancer

  2. dys genomic structure FTF Q72L FTF T31N Exon1 2 3 4 5 6 7 8 9 10 LP05454 3.8kb F2F elution 1.7kb 2.7kb 3.8kb 3.8kb

  3. CG13196 enhancer

  4. Expression patterns of CG13196 enhancer and TCGTG enhancer 4xTCGTG GFP Dys CG13196 GFP Dys Early S15 S16

  5. Ectopic expression of dys can activate CG13196 and TCGTG enhancer ectopically CG13196 Dys 4xTCGTG GFP Dys S15 S16

  6. CG15252 5’ upstream TCGTG TCGTG TCGTG Exon1 3.2kb 0.8 kb TCGTG cluster 9.0 kB 5’ upstream

  7. Spalt is not required for CG15252 activation by dys in DT CG15252 Dys CG15252 Dys btl-gal4;UAS-dys btl-gal4;UAS-spalt;UAS-dys Need to do: CG15252 expression in Spalt mutants

  8. a-Dys mbo-lacZ btl-gal4, UAS-dys Ectopic dys can activate mbo-lacZ ectopically

  9. mbo 3’ TCGTG ACGTG 407bp

  10. Conclusion • dys target genes can only be turn on at late stages (late 14 or early 15) • There is no transcriptional inhibition at early stage • There are other transcriptional activators turns on at late stages or changes in chromatin structure. • Spalt is not required for CG15252 expression

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