A conserved genetic interaction between Spt6 and Set2 regulates H3K36 methylation. Issue 8 (22nd February 2019)

• Record Type: Journal Article
• Authors: Gopalakrishnan, Rajaraman; Marr, Sharon K; Kingston, Robert E; Winston, Fred
• Is Part Of: Nucleic acids research. Volume 47:Issue 8(2019)
• Page Start: 3888
• Page End: 3903
• Publication Date: 2019-02-22
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkz119
• Languages: English
• ISSNs: 0305-1048

Abstract

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 ( SET2 sup mutations) in a region encoding a proposed autoinhibitory domain. SET2 sup mutations suppress the H3K36 methylation defect in the spt6 mutant, as well as in other mutants that impair H3K36 methylation. We also show that SET2 sup mutations overcome the requirement for certain Set2 domains for H3K36 methylation. In vivo, SET2 sup mutants have elevated levels of H3K36 methylation and the purified Set2 sup mutant protein has greater enzymatic activity in vitro . ChIP-seq studies demonstrate that the H3K36 methylation defect in the spt6 mutant, as well as its suppression by a SET2 sup 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 autoinhibitory domain requires multiple Set2 interactions to ensure that H3K36 methylation occurs specifically on actively transcribed chromatin.