A conserved genetic interaction between Spt6 and Set2 regulates H3K36 methylation

Nucleic Acids Research
Rajaraman GopalakrishnanFred Winston

Abstract

The transcription elongation factor Spt6 and the H3K36 methyltransferase Set2 are both required for H3K36 methylation and transcriptional fidelity in Saccharomyces cerevisiae. However, the nature of the requirement for Spt6 has remained elusive. By selecting for suppressors of a transcriptional defect in an spt6 mutant, we have isolated several highly clustered, dominant SET2 mutations (SET2sup mutations) in a region encoding a proposed autoinhibitory domain. SET2sup mutations suppress the H3K36 methylation defect in the spt6 mutant, as well as in other mutants that impair H3K36 methylation. We also show that SET2sup mutations overcome the requirement for certain Set2 domains for H3K36 methylation. In vivo, SET2sup mutants have elevated levels of H3K36 methylation and the purified Set2sup mutant protein has greater enzymatic activityin vitro. ChIP-seq studies demonstrate that the H3K36 methylation defect in the spt6 mutant, as well as its suppression by a SET2sup mutation, occurs at a step following the recruitment of Set2 to chromatin. Other experiments show that a similar genetic relationship between Spt6 and Set2 exists in Schizosaccharomyces pombe. Taken together, our results suggest a conserved mechanism by which the Set2 ...Continue Reading

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Citations

Sep 11, 2019·Current Protocols in Molecular Biology·Ameet ShettyFred Winston
Sep 11, 2019·Current Protocols in Molecular Biology·Rajaraman Gopalakrishnan, Fred Winston
Jul 2, 2020·G3 : Genes - Genomes - Genetics·Mitchell A EllisonKaren M Arndt
Apr 24, 2021·Genes & Development·Olga ViktorovskayaFred Winston
Jul 6, 2021·The Journal of Biological Chemistry·Ryan J Separovich, Marc R Wilkins

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Datasets Mentioned

BETA
GSE116646

Methods Mentioned

BETA
Hi-Seq
electrophoresis
PCR
transfection
immunoprecipitation
ChIP-Seq
ChIP
two-hybrid
co-immunoprecipitation

Software Mentioned

R scripts
Snakemake workflow manager
custom
Bowtie2
SPP package
samtools
Uniprot
bedtools
Licor ImageStudio
igvtools

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