Spt3 and Spt8 Are Involved in the Formation of a Silencing Boundary by Interacting with TATA-Binding Protein

Biomolecules. 2023 Mar 30;13(4):619. doi: 10.3390/biom13040619.

Abstract

In Saccharomyces cerevisiae, a heterochromatin-like chromatin structure called the silencing region is present at the telomere as a complex of Sir2, Sir3, and Sir4. Although spreading of the silencing region is blocked by histone acetylase-mediated boundary formation, the details of the factors and mechanisms involved in the spread and formation of the boundary at each telomere are unknown. Here, we show that Spt3 and Spt8 block the spread of the silencing regions. Spt3 and Spt8 are members of the Spt-Ada-Gcn5-acetyltransferase (SAGA) complex, which has histone acetyltransferase activity. We performed microarray analysis of the transcriptome of spt3Δ and spt8Δ strains and RT-qPCR analysis of the transcript levels of genes from the subtelomeric region in mutants in which the interaction of Spt3 with TATA-binding protein (TBP) is altered. The results not only indicated that both Spt3 and Spt8 are involved in TBP-mediated boundary formation on the right arm of chromosome III, but also that boundary formation in this region is DNA sequence independent. Although both Spt3 and Spt8 interact with TBP, Spt3 had a greater effect on genome-wide transcription. Mutant analysis showed that the interaction between Spt3 and TBP plays an important role in the boundary formation.

Keywords: SAGA complex; Saccharomyces cerevisiae; TBP; boundary; gene silencing; subtelomere.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Histone Acetyltransferases / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins* / genetics
  • Saccharomyces cerevisiae Proteins* / metabolism
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae / genetics
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae / metabolism
  • TATA-Box Binding Protein / genetics
  • TATA-Box Binding Protein / metabolism
  • Transcription Factors* / metabolism

Substances

  • Transcription Factors
  • TATA-Box Binding Protein
  • Saccharomyces cerevisiae Proteins
  • Histone Acetyltransferases
  • SPT3 protein, S cerevisiae
  • SIR3 protein, S cerevisiae
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae
  • SPT8 protein, S cerevisiae

Grants and funding

This work was supported by JSPS KAKENHI Grant Number JP 22H02301.