SUMOylation of Bonus, the Drosophila homolog of Transcription Intermediary Factor 1, safeguards germline identity by recruiting repressive chromatin complexes to silence tissue-specific genes
- California Institute of Technology, Division of Biology and Biological Engineering, United States
- Institute of Gene Biology, Russian Academy of Sciences, Russian Federation
- University of California, Riverside, United States
Figures
Figure 1 with 1 supplement
Germline expression of Bonus is required for oogenesis.
(A) Bon is expressed throughout oogenesis. Stacked confocal image of wild-type Oregon-R flies stained for Bon. (B) Bar graph shows the relative expression of Bon (normalized to rp49 level) in …
Figure 1—figure supplement 1
An important function of Bonus in the early stages of oogenesis.
(A) Schematics of a Drosophila ovariole and the expression pattern of different Gal4 driver lines during developmental stages of oogenesis. (B) TUNEL assay. Confocal images of egg chambers from …
Figure 2 with 1 supplement
Bonus functions as a repressor of tissue-specific genes in ovary.
(A) Bon GLKD leads to misexpression of tissue-specific genes in the ovary. Volcano plot shows fold changes in genes expression upon Bon GLKD driven by nos-Gal4 in the ovary as determined by RNA-seq …
Figure 2—figure supplement 1
Depletion of Bon induces ectopic activation of non-ovarian genes.
(A) Scatterplots display changes in transposons expression upon Bon GLKD driven by MT-Gal4 (left) or nos-Gal4 (right) as determined by RNA-seq data (log2-transformed TPM values, n = 3). Flies with wh…
Figure 3 with 1 supplement
Bonus induces transcriptional silencing.
(A) Schematics of the reporter construct in flies that allows Bon recruitment to nascent reporter transcript in flies. λN-GFP-Bonus and the mKate reporter encoding 4BoxB hairpins are co-expressed in …
Figure 3—figure supplement 1
Bonus tethering leads to transcriptional silencing of the reporter.
(A) Bar plot shows the Luciferase reporter expression (normalized to rp49 level) in ovaries upon tethering of λN-GFP-Bonus or λN-GFP control ovaries (RT-qPCR, dots correspond to three independent …
Figure 4 with 1 supplement
Bonus interacts with Mi-2, Rpd3, and SetDB1.
(A) Reporter silencing by Bon depends on Mi-2 and SetDB1. Bar plot showing the reporter expression (normalized to rp49 level) upon tethering of control λN-GFP or λN-GFP-Bonus in ovaries with Rpd3, …
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Figure 4—source data 1
Annotated and uncropped western blots and raw images for Figure 4B.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig4-data1-v1.zip
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Figure 4—source data 2
Annotated and uncropped western blots and raw images for Figure 4C.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig4-data2-v1.zip
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Figure 4—source data 3
Annotated and uncropped western blots and raw images for Figure 4D.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig4-data3-v1.zip
Figure 4—figure supplement 1
Bonus interacts with Mi-2.
Western blot analysis of immunoprecipitation experiment using GFP nanotrap beads from S2 cells co-expressing GFP-Bonus and Flag-tagged Mi-2 (A) or Flag-tagged Mep-1 (B). Lysates not expressing the …
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Figure 4—figure supplement 1—source data 1
Annotated and uncropped western blots and raw images for Figure 4—figure supplement 1A.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig4-figsupp1-data1-v1.zip
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Figure 4—figure supplement 1—source data 2
Annotated and uncropped western blots and raw images for Figure 4—figure supplement 1B.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig4-figsupp1-data2-v1.zip
Figure 5 with 2 supplements
Bonus is SUMOylated.
(A) Schematic representation of putative SUMOylation sites within Bon. SUMOylation consensus sites are shown and boxed. Canonical consensus sites are in bold. Putative SUMOylated lysines were …
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Figure 5—source data 1
Annotated and uncropped western blots and raw images for Figure 5B.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-data1-v1.zip
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Figure 5—source data 2
Annotated and uncropped western blots and raw images for Figure 5C.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-data2-v1.zip
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Figure 5—source data 3
Annotated and uncropped western blots and raw images for Figure 5F.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-data3-v1.zip
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Figure 5—source data 4
Annotated and uncropped western blots and raw images for Figure 5I.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-data4-v1.zip
Figure 5—figure supplement 1
SUMOylation of Bonus.
(A) Western blot analysis of GFP-Bonus SUMOylation in S2 cells (top) and ovaries (bottom). Total protein lysates from S2 cells co-expressing HA-SUMO and GFP-Bonus or from ovaries of flies …
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Figure 5—figure supplement 1—source data 1
Annotated and uncropped western blots and raw images for Figure 5—figure supplement 1A.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-figsupp1-data1-v1.zip
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Figure 5—figure supplement 1—source data 2
Annotated and uncropped western blots and raw images for Figure 5—figure supplement 1B.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-figsupp1-data2-v1.zip
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Figure 5—figure supplement 1—source data 3
Annotated and uncropped western blots and raw images for Figure 5—figure supplement 1C.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-figsupp1-data3-v1.zip
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Figure 5—figure supplement 1—source data 4
Annotated and uncropped western blots and raw images for Figure 5—figure supplement 1D.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-figsupp1-data4-v1.zip
Figure 5—figure supplement 2
SUMO-independent and dependent interactions of Bonus.
(A, B) Western blot analysis shows the SUMO-independent interaction between Bon and Mi-2 or Rpd3. Total protein lysates from S2 cells co-expressing GFP-Bonus (WT) or triple mutant GFP-Bonus[3KR] …
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Figure 5—figure supplement 2—source data 1
Annotated and uncropped western blots and raw images for Figure 5—figure supplement 2A.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-figsupp2-data1-v1.zip
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Figure 5—figure supplement 2—source data 2
Annotated and uncropped western blots and raw images for Figure 5—figure supplement 2B.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-figsupp2-data2-v1.zip
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Figure 5—figure supplement 2—source data 3
Annotated and uncropped western blots and raw images for Figure 5—figure supplement 2C.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig5-figsupp2-data3-v1.zip
Figure 6 with 1 supplement
SUMO E3-ligase Su(var)2–10 interacts with Bonus and regulates its SUMOylation.
(A) Western blot analysis shows the interaction between Bon and SUMO E2-conjugating enzyme Ubc9. Total protein lysates from S2 cells co-expressing Flag-Ubc9 and GFP-Bonus (WT), SUMO-deficient triple …
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Figure 6—source data 1
Annotated and uncropped western blots and raw images for Figure 6A.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig6-data1-v1.zip
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Figure 6—source data 2
Annotated and uncropped western blots and raw images for Figure 6B.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig6-data2-v1.zip
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Figure 6—source data 3
Annotated and uncropped western blots and raw images for Figure 6C.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig6-data3-v1.zip
Figure 6—figure supplement 1
Bonus does not interact with Ubc9 in S2 cells.
Western blot analysis of immunoprecipitation experiment using GFP nanotrap beads from S2 cells co-expressing Flag-tagged Ubc9 and GFP-Bonus (A) or Flag-tagged Ubc9 and GFP-Bonus or SUMO-deficient …
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Figure 6—figure supplement 1—source data 1
Annotated and uncropped western blots and raw images for Figure 6—figure supplement 1A.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig6-figsupp1-data1-v1.zip
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Figure 6—figure supplement 1—source data 2
Annotated and uncropped western blots and raw images for Figure 6—figure supplement 1B.
- https://cdn.elifesciences.org/articles/89493/elife-89493-fig6-figsupp1-data2-v1.zip
