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. 2018 Jan 16;22(3):611-623.
doi: 10.1016/j.celrep.2017.12.079.

INO80 Chromatin Remodeling Coordinates Metabolic Homeostasis with Cell Division

Graeme J Gowans  1 , Alicia N Schep  2 , Ka Man Wong  1 , Devin A King  1 , William J Greenleaf  2 , Ashby J Morrison  3
Affiliations

INO80 Chromatin Remodeling Coordinates Metabolic Homeostasis with Cell Division

Graeme J Gowans et al. Cell Rep. .

Abstract

Adaptive survival requires the coordination of nutrient availability with expenditure of cellular resources. For example, in nutrient-limited environments, 50% of all S. cerevisiae genes synchronize and exhibit periodic bursts of expression in coordination with respiration and cell division in the yeast metabolic cycle (YMC). Despite the importance of metabolic and proliferative synchrony, the majority of YMC regulators are currently unknown. Here, we demonstrate that the INO80 chromatin-remodeling complex is required to coordinate respiration and cell division with periodic gene expression. Specifically, INO80 mutants have severe defects in oxygen consumption and promiscuous cell division that is no longer coupled with metabolic status. In mutant cells, chromatin accessibility of periodic genes, including TORC1-responsive genes, is relatively static, concomitant with severely attenuated gene expression. Collectively, these results reveal that the INO80 complex mediates metabolic signaling to chromatin to restrict proliferation to metabolically optimal states.

Keywords: Abf1; Arp5; INO80; Msn2/4; Reb1; TORC1; cell division; chromatin; metabolism; yeast metabolic cycle.

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Conflict of interest statement

DECLARATION OF INTERESTS

The authors declare no competing interests.

Figures

Figure 1
Figure 1. The INO80 Complex Is Essential for Respiration Oscillations
(A) Organization of the yeast metabolic cycle (YMC). Top: respiration cycles are shown with the corresponding percentage of dissolved oxygen (dO2) in culture. Bottom: relative fold change gene expression of previously classified periodic genes (Tu et al., 2005). RC, reductive charging (1,508 periodic genes); OX, oxidative (1,016 periodic genes); RB, reductive building (975 periodic genes). (B) dO2 traces in the wild-type and indicated mutant strains lacking subunits of the INO80 complex. (C) Illustration of the INO80 complex with structural modules noted fromTosi et al. (2013). Colors denote the severity of YMC defects observed upon deletion of the indicated subunit. See also Figure S1 and Table S1.
Figure 2
Figure 2. Disruption of the INO80 Complex Alters Global Transcription across the YMC
(A) Left: YMC dO2 traces in the wild-type and arp5Δ mutant. Samples were taken at the indicated time points (coded by color) for RNA-seq and PCA. Right: principal components (PCs) 1 and 2 are shown. Technical replicates are displayed for each time point and separated by strain. (B) Left: mean expression of periodic genes in wild-type (WT) and arp5Δ strains within each YMC phase. n = number of periodic genes in each phase previously classified (Kuang et al., 2014). Expression of each gene is shown as FPKM. Right: expression of each individual gene is shown as heatmaps (scaled by row). (C) DAVID functional enrichment analysis of significantly differentially expressed (SDE) genes at comparable time points between the wild-type and arp5Δ mutant (refer to Experimental Procedures for more detail). Bar colors denote the wild-type phase with which periodic genes are associated. Enrichment scores are −log10(p value). See also Table S2.
Figure 3
Figure 3. Cell Division Is Disconnected from the YMC in the arp5Δ Mutant
(A) k-means clustering of previously classified periodic genes (Kuang et al., 2014) following z-score normalization of RNA-seq data for each gene, time point, and strain. (B) DAVID functional enrichment of genes in the arp5Δ mutant that do not cluster in the corresponding YMC phase. Annotations related to cell cycle are highlighted. APC, anaphase-promoting complex. Enrichment scores are −log10(p value). (C) Samples were taken at indicated time points across wild-type and arp5Δ YMCs. Top: dO2 traces and ratio of G2 to G1 cells for each strain. Bottom: representative DNA content histograms from samples indicated with a thicker outline at the top. (D) Number of budding cells, indicative of M phase entry, in samples across wild-type and arp5Δ YMCs. See also Table S3.
Figure 4
Figure 4. Loss of INO80 Function Alters Global Chromatin Accessibility in the YMC
(A) PCA plot of ATAC-seq data from samples taken at time points shown in Figure 2A. Replicates are displayed for each time point and strain (colored as in Figure 2A). (B) Comparison of mean ATAC-seq scores for periodic genes from each phase of wild-type and arp5Δ YMC following Z score normalization for each gene and time point. The RC, OX, and RB phases are highlighted. (C) Chromosomal plot of log2 mean wild-type and arp5Δ ATAC-seq and RNA-seq (FPKM) in 500 bp bins for the YMC time points described in Figure 2A. Scores in both wild-type and arp5Δ cells were normalized to mean wild-type. Red and blue boxes highlight static regions of increased and decreased accessibility, respectively, in arp5Δ cells. Gray boxes indicate the corresponding chromosomal regions in wild-type cells. See also Figures S2 and S3.
Figure 5
Figure 5. Metabolic Gene Promoters Are Dependent on INO80-Regulated Chromatin Accessibility
(A) Promoter ATAC accessibility scores organized by all annotated transcription factor (TF) motifs are shown for both wild-type and arp5Δ YMCs. (B) Magnification of indicated region in (A). The time points and YMC phase from Figure 2A are shown. (C) Mean RNA-seq FPKM for genes that contain the indicated transcription factor motifs in their promoters. (D) Left: illustration of the TORC1 pathway for the indicated transcription factors. Right: DAVID significant functional enrichment of genes with the indicated transcription factor motifs. SAM, S-adenosyl-methionine; TCA, tricarboxylic acid. (E) ATAC accessibility is shown as the number of Tn5 insertions per million reads around the indicated transcription factor motif for wild-type (green) and arp5Δ (pink) samples. (F) +1 and −1 nucleosome densities, relative to transcription start sites, containing Abf1 or Reb1 motifs. Wild-type (green) and arp5Δ (pink) samples across all time points of the YMC are shown. Red/blue dotted lines denote the median nucleosome position for the wild-type and arp5Δ, respectively. p = 0.002 for Abf1 and 0.158 for Reb1. See also Figure S4.
Figure 6
Figure 6. The INO80 Complex Is a Key Regulator of TORC1-Responsive Gene Expression
(A) Top: same as Figure 2A. YMC dO2 traces in the wild-type and arp5Δ mutant. RNA-seq samples were taken at the indicated time points. Mean expression is shown as FPKM of Msn2-induced genes (center, Msn2 genes) and rapamycin-sensitive genes (bottom, Rap genes) across the YMC. Right: gene expression heatmaps of rapamycin-sensitive genes scaled by row. (B) Pearson’s correlation between the RNA transcript levels across the wild-type YMC and those in an asynchronous culture before and after rapamycin treatment (30 nM) for the indicated time in minutes. (C) Western blot analysis of the indicated wild-type time points with an antibody that recognizes the phosphorylated TORC1 substrate, mammalian homolog ribosomal protein S6 (Rps6). Hexokinase 1 (Hxk1) is shown as a loading control. (D) Samples were taken across a wild-type YMC at the indicated time points before and after treatment with rapamycin (50 nM), indicated by the dashed line. Also shown is a western blot analysis using antibodies that recognize pRps6, Hxk1, acetylated histone H3 lysine 9, (H3K9ac), and histone H3 (H3). (E) Western blot analysis of the arp5Δ mutant YMC as in (C). See also Figure S5 and Table S4.
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