Single Cell Visualization of Yeast Gene Expression Shows Correlation of Epigenetic Switching between Multiple Heterochromatic Regions through Multiple Generations

Mano, Yasunobu; Kobayashi, Tetsuya; Nakayama, Jun‐ichi; Uchida, Hiroyuki; Oki, Masaya

doi:10.1371/journal.pbio.1001601

Cited by 28 publications

(43 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Colonies of yeast that are ade1 Δ are a dark red colour; reintroduction of functional Ade1 protein restores the wild type white coloration in a dose-dependent manner. Such differences in colony colour and ability to grown on adenine dropout media (SD-Ade) have been utilized to differentiate strains of yeast prions37, assess mitotic stability38 and monitor gene expression39.…”

Section: Resultsmentioning

confidence: 99%

Rapid generation of hypomorphic mutations

et al. 2017

View full text Add to dashboard Cite

Hypomorphic mutations are a valuable tool for both genetic analysis of gene function and for synthetic biology applications. However, current methods to generate hypomorphic mutations are limited to a specific organism, change gene expression unpredictably, or depend on changes in spatial-temporal expression of the targeted gene. Here we present a simple and predictable method to generate hypomorphic mutations in model organisms by targeting translation elongation. Adding consecutive adenosine nucleotides, so-called polyA tracks, to the gene coding sequence of interest will decrease translation elongation efficiency, and in all tested cell cultures and model organisms, this decreases mRNA stability and protein expression. We show that protein expression is adjustable independent of promoter strength and can be further modulated by changing sequence features of the polyA tracks. These characteristics make this method highly predictable and tractable for generation of programmable allelic series with a range of expression levels.

show abstract

Section: Resultsmentioning

confidence: 99%

Rapid generation of hypomorphic mutations

et al. 2017

View full text Add to dashboard Cite

show abstract

“…This question has been addressed with pairs of fluorescent reporter genes integrated at separate silenced locations. In one study, correlated switches in expression state were observed in cell lineages when the genes were placed at the edges of the silent chromatin domains at HMR and HML (Mano et al 2013). These results indicate that inheritance of silent chromatin is influenced by a property that is generic to the cell and not specific to the locus.…”

Section: Variegated Expression and Inheritancementioning

confidence: 98%

“…When reporters were placed at an artificial telomere or beyond the silencers at HMR expression, their expression was variegated (Gottschling et al 1990;Simms et al 2008;Mano et al 2013). In the case of the ADE2 reporter, silenced cells acquired a red pigment whereas genetically identical, nonsilent cells remained white.…”

Section: Variegated Expression and Inheritancementioning

confidence: 99%

The Nuts and Bolts of Transcriptionally Silent Chromatin in Saccharomyces cerevisiae

2016

View full text Add to dashboard Cite

Transcriptional silencing in Saccharomyces cerevisiae occurs at several genomic sites including the silent mating-type loci, telomeres, and the ribosomal DNA (rDNA) tandem array. Epigenetic silencing at each of these domains is characterized by the absence of nearly all histone modifications, including most prominently the lack of histone H4 lysine 16 acetylation. In all cases, silencing requires Sir2, a highly-conserved NAD + -dependent histone deacetylase. At locations other than the rDNA, silencing also requires additional Sir proteins, Sir1, Sir3, and Sir4 that together form a repressive heterochromatin-like structure termed silent chromatin. The mechanisms of silent chromatin establishment, maintenance, and inheritance have been investigated extensively over the last 25 years, and these studies have revealed numerous paradigms for transcriptional repression, chromatin organization, and epigenetic gene regulation. Studies of Sir2-dependent silencing at the rDNA have also contributed to understanding the mechanisms for maintaining the stability of repetitive DNA and regulating replicative cell aging. The goal of this comprehensive review is to distill a wide array of biochemical, molecular genetic, cell biological, and genomics studies down to the "nuts and bolts" of silent chromatin and the processes that yield transcriptional silencing.

show abstract

“…For example, loss of the euchromatin-associated bromodomain protein Bdf1 or histone variant H2A.Z results in expanded heterochromatin regions [98, 99]. As a result of such competition, negotiable borders are associated with frequent changes of epigenetic states [100]. The classical example of PEV in Drosophila , where white is silenced in a portion of progenitor cells during early development, also suggests that heterochromatin spreads over varying distances rather than being constrained to a defined location.…”

Section: Negotiable Bordersmentioning

confidence: 99%

Chromosome boundary elements and regulation of heterochromatin spreading

Wang

Lawry

Cohen

et al. 2014

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

Chromatin is generally classified as euchromatin or heterochromatin, each with distinct histone modifications, compaction levels, and gene expression patterns. Although the proper formation of heterochromatin is essential for maintaining genome integrity and regulating gene expression, heterochromatin can also spread into neighboring regions in a sequence-independent manner, leading to the inactivation of genes. Because the distance of heterochromatin spreading is stochastic, the formation of boundaries, which block the spreading of heterochromatin, is critical for maintaining stable gene expression patterns. Here we review the current understanding of the mechanisms underlying heterochromatin spreading and boundary formation.

show abstract

Single Cell Visualization of Yeast Gene Expression Shows Correlation of Epigenetic Switching between Multiple Heterochromatic Regions through Multiple Generations

Abstract: A single-cell method allows the assessment of relationships between the dynamic epigenetic behavior of yeast heterochromatin boundaries over multiple generations.

Cited by 28 publications

References 39 publications

Rapid generation of hypomorphic mutations

Rapid generation of hypomorphic mutations

The Nuts and Bolts of Transcriptionally Silent Chromatin in Saccharomyces cerevisiae

Chromosome boundary elements and regulation of heterochromatin spreading

Contact Info

Product

Resources

About