Study / S129MPFB72021-08-09

Cis-regulatory variants affect gene expression dynamics in yeast

Ching-Hua Shih, Justin Fay

About this study

Evolution of cis-regulatory sequences depends on how they affect gene expression and motivates both the identification and prediction of cis-regulatory variants responsible for expression differences within and between species. While much progress has been made in relating cis-regulatory variants to expression levels, the timing of gene activation and repression may also be important to the evolution of cis-regulatory sequences. We investigated allele-specific expression (ASE) dynamics within and between Saccharomyces species during the diauxic shift and found appreciable cis-acting variation in gene expression dynamics. Within-species ASE is associated with intergenic variants, and ASE dynamics are more strongly associated with insertions and deletions than ASE levels. To refine these associations, we used a high-throughput reporter assay to test promoter regions and individual variants. Within the subset of regions that recapitulated endogenous expression, we identified and characterized cis-regulatory variants that affect expression dynamics. Between species, chimeric promoter regions generate novel patterns and indicate constraints on the evolution of gene expression dynamics. We conclude that changes in cis-regulatory sequences can tune gene expression dynamics and that the interplay between expression dynamics and other aspects of expression is relevant to the evolution of cis-regulatory sequences.

Full author list & citation

Ching-Hua Shih, Justin Fay. Cis-regulatory variants affect gene expression dynamics in yeast. 2021-08-09. https://doi.org/10.7554/eLife.68469

Experiments 3

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Intra-specific Oak × ChII CRE-seq time course

An integrated CRE-seq library tested 130-bp promoter windows, parental alleles, and single-variant substitutions from the Oak (YJF153) and China II (SX6) Saccharomyces cerevisiae strains in an Oak × ChII diploid. Four random 10-bp barcodes were designed per construct, and barcode RNA/DNA activity was measured over 19 diauxic-shift timepoints.

Targeted Genomic Integration MPRABudding yeastS288c R64-1-1
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Inter-specific S. cerevisiae × S. uvarum CRE-seq time course

An integrated CRE-seq library tested 130-bp promoter windows from S. cerevisiae and S. uvarum, including both parental alleles and reciprocal distal/proximal chimeras, in an Oak × S. uvarum diploid. Four random 10-bp barcodes were designed per construct, and barcode RNA/DNA activity was measured over 27 timepoints.

Targeted Genomic Integration MPRABudding yeastS288c R64-1-1
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Full-length Oak/ChII promoter CRE-seq validation

Full-length promoters from ALD5, GND2, and PHO3 in Oak and ChII backgrounds were barcoded and integrated at URA3 in the Oak × ChII diploid. The supplied full-length count/activity file was summarized by gene, allele, and measured timepoint.

Targeted Genomic Integration MPRABudding yeastS288c R64-1-1
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Raw source data 18 files

Original supplemental and deposited inputs retained for this study. Download files individually or together as a ZIP; nested folders are preserved. Source reuse terms apply, and sequencing reads may be omitted.

Download all 18 files (ZIP)CRE.YJF1455.type.csvCRE.YJF1484.type.csvCRE_analysis.RCRE_Libraries.YJF1455.csvCRE_Libraries.YJF1484.csvfull.length.counts.csvinter.counts.csvinter.stats.csvintra.counts.csvintra.stats.csvmainTableR3.YJF1455.csvmainTableR3.YJF1484.csvREADME_file_descriptions.pdfweighted_dwtest.RYJF1455.annotall.csvYJF1455.ASE.csvYJF1455.genes.csvYJF1484.ASE.csv

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