fusion cell gene profiling identify mbo and cg15252 enhancer molecular mechanism of target gene...

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

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Page 1: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

•Fusion cell gene profiling

•Identify mbo and CG15252 enhancer

•Molecular mechanism of target gene regulation by Dys::Tgo

•Dys::Tgo activation of CG13196

•Dys:Tgo activation of CG15252

•Identify dys fusion cell enhancer

dys-gal4::UAS-CD8-GFP btl-gal4::UAS-CD8-GFP

dys-gal4::UAS-CD8-GFP dys-gal4::UAS-CD8-GFP ; dys/dys

•Identify dys target gene

Page 2: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

dys genomic structure

FTF Q72L FTF T31N

F2F elution

LP05454

Exon1 2 3 4 5 6 7 8 9 10

1.7kb2.7kb

3.8kb

3.8kb

3.8kb

Page 3: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

CG13196 enhancer

Page 4: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

Expression patterns of CG13196 enhancer and TCGTG enhancer

Early S15

4xTCGTG GFP Dys

S16

CG13196 GFP Dys

Page 5: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

CG13196 4xTCGTG GFP

S15

S16

Dys Dys

Ectopic expression of dys can activate CG13196 and TCGTG enhancer ectopically

Page 6: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

CG15252 5’ upstream

Exon1

3.2kb

9.0 kB 5’ upstream

0.8 kb TCGTG cluster

TCGTG TCGTG TCGTG

Page 7: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

Spalt is not required for CG15252 activation by dys in DT

Need to do:CG15252 expression in Spalt mutants

CG15252 Dys CG15252 Dys

btl-gal4;UAS-dys btl-gal4;UAS-spalt;UAS-dys

Page 8: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

Ectopic dys can activate mbo-lacZ ectopically

mbo-lacZ-Dys

btl-gal4, UAS-dys

Page 9: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

mbo 3’

TCGTG

407bp

ACGTG

Page 10: Fusion cell gene profiling Identify mbo and CG15252 enhancer Molecular mechanism of target gene regulation by Dys::Tgo Dys::Tgo activation of CG13196 Dys:Tgo

Conclusion

•There are other transcriptional activators turns on at late stages or changes in chromatin structure.

• Spalt is not required for CG15252 expression

•There is no transcriptional inhibition at early stage

• dys target genes can only be turn on at late stages (late 14 or early 15)