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Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae

ABSTRACT: Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae

PROVIDER: PRJNA785980 | ENA |

REPOSITORIES: ENA

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Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae [I]

Project description:Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae [I]

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Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae [II]

Project description:Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae [II]

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Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae [I]

Project description:The formation of heterochromatin at HML, HMR, and telomeres in Saccharomyces cerevisiae involves two main steps: Recruitment of Sir proteins to silencers and their spread throughout the silenced domain. For the following datasets, we created a fusion protein between the heterochromatin protein Sir3 and the non-site-specific bacterial adenine methyltransferase M.EcoGII, with or without a 3xV5 epitope at the C-terminus. We performed ChIP-seq experiments (immunoprecipitated Sir3-M.EcoGII-3xV5) and MeDIP-seq experiments (immunoprecipitated m6A methylated DNA). We also used a temperature-sensitive allele of SIR3 (sir3-8) fused to M.ECOGII to induce m6A methylation for MeDIP-seq.

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Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae [II]

Project description:The formation of heterochromatin at HML, HMR, and telomeres in Saccharomyces cerevisiae involves two main steps: Recruitment of Sir proteins to silencers and their spread throughout the silenced domain. For the following datasets, we created a fusion protein between the heterochromatin protein Sir3 and the non-site-specific bacterial adenine methyltransferase M.EcoGII. We mapped sites of Sir3-chromatin interactions genome-wide using long-read Nanopore sequencing to detect adenines methylated by the fusion protein. We also used a temperature-sensitive allele of SIR3 (sir3-8) fused to M.ECOGII to induce m6A methylation over time. Time courses involved a switch from restrictive temperature (37°C) to permissive temperature (25°C).

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Distinguishing between recruitment and spread of silent chromatin structures in Saccharomyces cerevisiae

Project description:This SuperSeries is composed of the SubSeries listed below.

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Project description:Flagellins from commensal bacteria can be weak Toll-like receptor (TLR)5 agonists despite high affinity binding to TLR5, ligands we termed “silent flagellins”. To determine if silent flagellins are detectable in the human gut, endogenous flagellins produced by the microbiota were isolated from stool obtained from a healthy adult female donor. TLR5 was used as bait to enrich for silent flagellins and TLR5-bound flagellins were identified by searching peptides against a custom flagellin database built from metagenome sequences.

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