piRNA-guided co-transcriptional silencing coopts nuclear export factors
- University of Cambridge, United Kingdom
Figures
Figure 1 with 2 supplements
Nxf2 is a piRNA pathway factor that functions in transcriptional gene silencing.
(A) Volcano plot showing enrichment values and corresponding significance levels for proteins co-purified with GFP-Panx from ovary lysates (n = 4 for GFP-Panx and n = 3 for control ovaries). …
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Figure 1—source data 1
List of proteins recovered in GFP-Panx IP-MS.
- https://doi.org/10.7554/eLife.47999.006
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Figure 1—source data 2
List of proteins recovered in GFP-Nxf2 IP-MS.
- https://doi.org/10.7554/eLife.47999.007
Figure 1—figure supplement 1
Nxf2 is a piRNA pathway factor acting in TGS.
(A) Western blot analyses of GFP-trap co-immunoprecipitation from lysates of ovaries from the indicated genotypes (representative replicate subjected to mass spectrometry analysis; IN, input; UB, …
Figure 1—figure supplement 2
Loss of Nxf2 impairs TGS at levels comparable to Panx.
(A) Box plot showing changes in H3K9me3 levels between nxf2 heterozygotes and mutants in ovarian ChIP-seq for the indicated categories (coloring of transposons is based on their fold change in …
Figure 2 with 1 supplement
Protein stability and localization of Nxf2 and Panx is reciprocally co-dependent.
(A) Expression and localization of Piwi and GFP-Nxf2 in egg chambers upon germline-specific knockdown (GLKD) of the indicated factors are shown by immunofluorescence (GFP, green; Piwi, magenta; DNA, …
Figure 2—figure supplement 1
Protein stability and localization of Nxf2 and Panx is reciprocally co-dependent.
(A) Expression and localization of Piwi and GFP-Nxf2 in egg chambers upon follicle cell-specific knockdown (tjKD) of the indicated factors are shown by immunofluorescence (GFP, green; Piwi, magenta; …
Figure 3 with 2 supplements
Requirements for the formation and function of the PICTS complex.
(A) Cartoons displaying the Nxf2 and Panx protein structures and domain mutants used (see also Figure 3—figure supplement 1A and Figure 3—figure supplement 2A). NTR, amino-terminal region; LRR, …
Figure 3—figure supplement 1
Interaction between Nxf2 and Panx is required for proper transposon silencing.
(A) Cartoons displaying the Panx protein structure and domain mutants used in panels (C) and (D). CC, coiled coil domain; HA, Hemagglutinin tag. (B) Representative western blot displaying …
Figure 3—figure supplement 2
Interaction between Nxf2 and Panx is required for proper transposon silencing.
(A) Cartoons displaying the Nxf2 protein structure and domain mutants and altered residues used in panels (B) through (H). (B) Western blot analyses of FLAG-tag co-immunoprecipitation from lysates …
Figure 4 with 1 supplement
Recruitment of PICTS components to nascent RNA results in chromatin silencing.
(A) Cartoon displaying the RNA tethering sensor construct used in OSCs. The construct was stably integrated and features the Drosophila simulans ubiquitin promoter (which contains an intron), the …
Figure 4—figure supplement 1
Artificial recruitment of Nxf2 to RNA or DNA results in chromatin silencing.
(A) Representative western blot analyses of λN-3xFLAG-fusion proteins, the HA-ZsGreen sensor and a loading control (His3) from lysates of OSCs transfected with the indicated constructs, used for …
Additional files
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Source code 1
Bash script for counting transposon derived reads.
- https://doi.org/10.7554/eLife.47999.015
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Source code 2
R code for differential expression analysis.
- https://doi.org/10.7554/eLife.47999.016
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Supplementary file 1
Oligonucleotides used in this study.
- https://doi.org/10.7554/eLife.47999.017
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Transparent reporting form
- https://doi.org/10.7554/eLife.47999.018
