S-phase-independent silencing establishment in Saccharomyces cerevisiae.

ELife
Davis Goodnight, Jasper Rine

Abstract

The establishment of silent chromatin, a heterochromatin-like structure at HML and HMR in Saccharomyces cerevisiae, depends on progression through S phase of the cell cycle, but the molecular nature of this requirement has remained elusive despite intensive study. Using high-resolution chromatin immunoprecipitation and single-molecule RNA analysis, we found that silencing establishment proceeded via gradual repression of transcription in individual cells over several cell cycles, and that the cell-cycle-regulated step was downstream of Sir protein recruitment. In contrast to prior results, HML and HMR had identical cell-cycle requirements for silencing establishment, with no apparent contribution from a tRNA gene adjacent to HMR. We identified the cause of the S-phase requirement for silencing establishment: removal of transcription-favoring histone modifications deposited by Dot1, Sas2, and Rtt109. These results revealed that silencing establishment was absolutely dependent on the cell-cycle-regulated interplay between euchromatic and heterochromatic histone modifications.

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Citations

Jan 23, 2021·Science·Marco Igor Valencia-SánchezKarim-Jean Armache

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Datasets Mentioned

BETA
GSE150737

Methods Mentioned

BETA
immunoprecipitation
ChIP
ChIP-seq
acetylation
flow cytometry
ubiquitination
PCR
immunoprecipitations
flow-cytometry

Software Mentioned

FISH
FIJI
custom Python scripts
Bowtie2
ggplot2
SAMtools
Micro
manager
IGV

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