Proteomic profiling of yeast heterochromatin connects direct physical and genetic interactions
Heterochromatin domains are stably repressed chromatin structures composed of a core assembly of silencing proteins that condense adjacent nucleosomes. The minimal heterochromatin structure can serve as a platform for recruitment of complementary regulatory factors. We find that a reconstituted budding yeast heterochromatin domain can act as a platform to recruit multiple factors that play a role in regulating heterochromatin function. We uncover the direct interaction between the SIR heterochromatin complex and a chromosomal boundary protein that restricts the spread of heterochromatin. We find that the SIR complex relieves a mechanism of auto-inhibition within the boundary protein Yta7, allowing the Yta7 bromodomain to engage chromatin. Our results suggest that budding yeast shares with other eukaryotes the ability to establish complex heterochromatin domains that coordinate multiple mechanisms of silencing regulation through physical interactions.
KeywordsHeterochromatin SIR complex Proteasome Yta7
We thank Danesh Moazed for initial support on this project. Thanks to Cassie Smith for preliminary test purifications of Yta7. This work was supported by NIH Grants T32GM008730 (A. Z.); R35GM119575 and K99/R00GM094291 (A. M. J.); NIH HG3456 and GM67945 (S. G.).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Gradolatto A, Smart SK, Byrum S, Blair LP, Rogers RS, Kolar EA, Lavender H, Larson SK, Aitchison JD, Taverna SD, Tackett AJ (2009) A noncanonical bromodomain in the AAA ATPase protein Yta7 directs chromosomal positioning and barrier chromatin activity. Mol Cell Biol 29:4604–4611CrossRefGoogle Scholar
- Krogan NJ, Dover J, Wood A, Schneider J, Heidt J, Boateng MA, Dean K, Ryan OW, Golshani A, Johnston M, Greenblatt JF, Shilatifard A (2003) The Paf1 complex is required for histone H3 methylation by COMPASS and Dot1p: linking transcriptional elongation to histone methylation. Mol Cell 11:721–729CrossRefGoogle Scholar
- Leggett DS, Glickman MH, Finley D (2005) Purification of proteasomes, proteasome subcomplexes, and proteasome-associated proteins from budding yeast. Methods Mol Biol 301:57–70Google Scholar
- Perez-Romero CA, Lalonde M, Chartrand P, Cusanelli E (2018) Induction and relocalization of telomeric repeat-containing RNAs during diauxic shift in budding yeast. Curr Genet 22:1Google Scholar