Transcription dynamically patterns the meiotic chromosome-axis interface
- New York University, United States
- University of Vienna, Austria
- Whitehead Institute for Biomedical Research, United States
Figures
Figure 1 with 1 supplement
Red1 preferentially localizes to the 3′ ends of convergent genes.
(A) Red1 distribution was plotted as an average across all genes on each of the 16 chromosomes as well as for the whole genome. The coding regions of genes were normalized to lengths of 1 kb (500 …
Figure 1—figure supplement 1
Genome-wide localization of meiotic axis proteins and cohesin.
(A) Distribution of axis proteins and cohesins as determined by ChIP-seq. Chromosome XI is shown as an example to show co-enrichment with the exception of the centromere (indicated by black circle). …
Figure 2
Red1 accumulates next to transcript ends of convergent gene pairs.
(A) Example of overlapping transcripts from a convergent gene pair. ORFs and transcripts of YKL077W and YKL075C were plotted with respect to their positions on chromosome XI. (B) Two-peak pattern of …
Figure 3 with 1 supplement
Transcriptional activity dictates Red1 binding patterns.
(A) Convergent gene pairs were identified, in which both genes were strongly transcribed (red) or both genes were weakly transcribed (blue) and average Red1 signal was determined around the …
Figure 3—figure supplement 1
Red1 position changes in response to transcription.
(A) Gene expression of the GAL2/SRL2 convergent gene pair in response to different concentrations of copper was measured by RT-qPCR at the indicated time points. Copper was added to the sporulation …
Figure 4 with 1 supplement
Transcriptional focusing depends on cohesin.
(A) Chromosomal localization of Red1 in rec8Δ mutants (blue) and WT (gray) on chromosomes XI and VI. Bottom panel shows zoom-in for a convergent gene pair on chromosome VI. (B) Red1 enrichment along …
Figure 4—figure supplement 1
Red1 binding in rec8Δ mutants.
(A) Distribution of widths of Red1 peaks in WT (green) and rec8Δ mutants (blue). (B) Probability of GAN motif occurrence around Red1 peak summits in WT (green) and rec8Δ mutants (blue). (C) Examples …
Figure 5 with 1 supplement
Chromosomal localization of Red1 requires Rec8 and Hop1 redundantly.
(A) Red1 binding profiles in WT (green), hop1Δ mutants (blue), and rec8Δ hop1Δ mutants (purple) from ChIP-seq experiments (NCIS normalized). A small (Chromosome I) and a medium sized chromosome …
Figure 5—figure supplement 1
Analysis of axis protein binding.
(A) ChIP-qPCR analysis of axis proteins at selected positions in the indicated mutant backgrounds was performed with the precipitated DNA of the respective ChIP-seq experiment, before library …
Figure 6
Red1 and Hop1 interact with Rec8.
(A) Illustration of the physical proximity assay M-track. The writer (Rec8) is tagged at on its C-terminus with a human derived histone lysine methyltransferase (HKMT), which will transfer up to …
Figure 7 with 1 supplement
Mitotic cohesin rescues transcriptional focusing but not genome-wide defects of rec8Δ mutants.
(A) Relative DSB hotspot intensity of each chromosome in WT (green) and rec8Δ mutants (blue) as determined using ssDNA arrays. Chromosomes are sorted by increasing size. (B) DSB activity spatially …
Figure 7—figure supplement 1
Ectopically expressed Scc1 localizes to the same sites as Rec8 in meiosis.
Top panel: ChIP–chip analysis of Rec8-3HA in otherwise WT cells (green). Bottom panel: ChIP–chip analysis of 3HA-Scc1 in rec8Δ, pREC8-3HA-SCC1 mutant cells (purple). Signal along chromosome XII is …
Figure 8
Model of the interface between axial elements and DNA.
Using the topological protein–DNA interaction of cohesin, a robust linkage between a protein rod (the axial element) and a transcriptionally active chromosome can be established. The protein axis …
Additional files
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Supplementary file 1
Strains and primers. (A) Strains used in the work. (B) Primers used for qPCR and RT-qPCR.
- https://doi.org/10.7554/eLife.07424.016
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Supplementary file 2
Python code for sequence extraction for motif analysis.
- https://doi.org/10.7554/eLife.07424.017
