Transcription of a 5’ extended mRNA isoform directs dynamic chromatin changes and interference of a downstream promoter
- The Francis Crick Institute, United Kingdom
- University of California, Berkeley, United States
Figures
Figure 1 with 1 supplement
NDC80luti transcription represses the NDC80ORF promoter.
(A) Expression pattern of two NDC80 mRNA isoforms during starvation and early meiosis. Diploid control cells (FW4644) or cells harboring the CUP1 promoter fused with IME1 and IME4 (pCUP-IME1/pCUP-IME…
Figure 1—figure supplement 1
NDC80luti transcription represses the NDC80ORF promoter.
(A) The same northern blot and western blot of NDC80 expression during a meiotic time course as described in Figure 1A, except that CIT1 and ribosomal RNA expression levels are also displayed. (B) …
Figure 2
Transcription of NDC80luti promotes H3K4me2 and H3K36me3 in the promoter and 5’ region of NDC80ORF.
(A) NDC80luti transcription promotes histone H3 lysine 4 dimethylation (H3K4me2) in the NDC80ORF promoter. Wild-type (FW1902) and set1Δ (FW3033) cells harboring the pCUP-IME1/pCUP-IME4 were induced …
Figure 3 with 1 supplement
Set2 and Set3 mediate NDC80luti induced gene repression of NDC80ORF.
(A) Set2 and Set3 are required for repression of NDC80ORF expression during early meiosis. Control (FW1902), set2Δ (FW2929), set3Δ (FW2928) and set2Δset3Δ (FW1922) cells harboring pCUP-IME1/pCUP-IME4…
Figure 3—figure supplement 1
The Set2 and the Set3 mediate NDC80luti induced gene repression of NDC80ORF.
(A) Quantification of NDC80ORF and NDC80luti levels in the experiment described in Figure 3A. Signals are normalized to SCR1. To control for variation in overall signal between different northern …
Figure 4 with 1 supplement
NDC80luti mediated repression is reversible.
(A) Schematic overview of the experimental set-up. (B) Cells repress NDC80luti and induce NDC80ORF when returned to a nutrient rich environment (YPD). Cells harboring ndt80Δ and pCUP-IME1/pCUP-IME4 …
Figure 4—figure supplement 1
NDC80luti mediated repression is reversible.
(A) Quantification of NDC80luti during return to rich medium as described in Figure 4B. RNA was extracted, reverse transcribed, and NDC80luti mRNA levels were determined by quantitative PCR. Signals …
Figure 5 with 1 supplement
Gene repression by NDC80luti is tunable.
(A) Adjustable expression of NDC80luti using the LexA-lexO system. Spot assay of cells harboring 0, 1, 2, 3, or 8 lexO binding sites in the NDC80luti promoter (UB8374, UB8358, UB8362, UB8366, …
Figure 5—figure supplement 1
Gene repression by NDC80luti is tunable.
(A) NDC80luti levels in the presence of variable number of lexO sites in the NDC80luti promoter. Cells harboring 0, 1, 2, 3, or 8 lexO and LexA-ER-AD (UB8374, UB8358, UB8362, UB8366, and UB8370) …
Figure 6
Model for NDC80luti mediated repression of NDC80ORF.
During mitosis the NDC80ORF promoter has an open chromatin structure and NDC80ORF is transcribed. During meiotic S-phase and meiotic prophase, transcription of NDC80luti deposits Set1-dependent …
Additional files
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Supplementary file 1
Table of yeast strains used in this study.
Table describing genotypes of the strains.
- https://doi.org/10.7554/eLife.27420.012
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Supplementary file 2
Table of oligo nucleotide sequences used in this study.
Table describing oligo nucleotide sequences used for quantitative PCR and for preparing the northern blot probes.
- https://doi.org/10.7554/eLife.27420.013
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Transparent reporting form
- https://doi.org/10.7554/eLife.27420.014
