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| Status |
Public on Sep 01, 2012 |
| Title |
Expression analyses in Drosophila young virgin female ovary |
| Organism |
Drosophila melanogaster |
| Experiment type |
Expression profiling by array
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| Summary |
Insulators delimit independent transcriptional domains within genomes by constraining enhancer and silencer action. These transcriptional effects depend upon DNA recognition by insulator binding proteins that recruit partners that protect against inappropriate long range modulation of non-target promoters. Insulator binding proteins are broadly expressed during development, with largely constitutive binding to thousands of genomic sites. Yet, tissue-specific transcriptional changes result from the loss of individual insulator binding proteins. To understand the molecular basis for such effects, we are studying the classic Drosophila insulator protein Suppressor of Hairy-wing [Su(Hw)]. Genetic studies show that loss of this broadly expressed insulator protein prevents oocyte development. To determine the basis for the block in oogenesis, we coupled transcriptional analyses in su(Hw) mutant ovaries with genome-wide definition of Su(Hw) binding in this tissue. These studies identified 71 direct targets of Su(Hw) regulation, with nearly 70% of these genes showing increased RNA accumulation when Su(Hw) is lost. Surprisingly, derepressed Su(Hw) target genes correspond to genes normally highly expressed in neural tissues, suggesting that Su(Hw) has a critical role in silencing neural genes in the ovary. Support for this postulate was obtained by genetic studies. We found that oocyte production was restored in su(Hw) mutant females that carry a deletion of one allele of the elav family RNA binding protein 9 (Rbp9) gene. These su(Hw) null oocytes can be fertilized, with evidence that embryos lacking Su(Hw) show compromised development. Our studies extend the known transcriptional activities of Su(Hw), indicating that Su(Hw) can function as an insulator, activator and repressor, the latter function being essential for oogenesis. These findings highlight that insulator proteins are versatile transcriptional regulatory proteins, suggesting that tissue specific contributions to transcription result from direct regulation of individual genes.
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| Overall design |
Ovaries for RNA isolation were dissected from 4-6 hour old virgin females of wild type, su(Hw)f/v (fertile), su(Hw)2/v (sterile) and su(Hw)Pb/2 (sterile) Drosophila melanogaster. At this stage of development, ovaries only contain egg chamber stages 1-8. Loss of Su(Hw) causes apoptosis at stage 9. Thus, the experimental design compares transcriptionl changes in the ovary prior to induction nof apoptosis in su(Hw) mutants.
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| Contributor(s) |
Soshnev AA, Geyer PK |
| Citation(s) |
23884443 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 GM042539 |
Regulation of Tissue Specific Gene Expression |
THE UNIVERSITY OF IOWA |
Pamela K Geyer |
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| Submission date |
Mar 15, 2012 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Alexey A Soshnev |
| E-mail(s) |
alexey.soshnev@gmail.com
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| Organization name |
University of Iowa
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| Department |
Biochemistry
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| Lab |
Geyer
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| Street address |
3159C MERF 375 Newton Rd
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| City |
Iowa City |
| State/province |
Iowa |
| ZIP/Postal code |
52246 |
| Country |
USA |
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| Platforms (1) |
| GPL1322 |
[Drosophila_2] Affymetrix Drosophila Genome 2.0 Array |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA153487 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE36528_RAW.tar |
26.0 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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