A synthetic non-histone substrate to study substrate targeting by the Gcn5 HAT and sirtuin HDACs

The Journal of Biological Chemistry
Anthony RösslMichael Downey

Abstract

Gcn5 and sirtuins are highly conserved histone acetyltransferase (HAT) and histone deacetylase (HDAC) enzymes that were first characterized as regulators of gene expression. Although histone tails are important substrates of these enzymes, they also target many nonhistone proteins that function in diverse biological processes. However, the mechanisms used by these enzymes to choose their nonhistone substrates are unknown. Previously, we used SILAC-based MS to identify novel nonhistone substrates of Gcn5 and sirtuins in yeast and found a shared target consensus sequence. Here, we use a synthetic biology approach to demonstrate that this consensus sequence can direct acetylation and deacetylation targeting by these enzymes in vivo Remarkably, fusion of the sequence to a nonsubstrate confers de novo acetylation that is regulated by both Gcn5 and sirtuins. We exploit this synthetic fusion substrate as a tool to define subunits of the Gcn5-containing SAGA and ADA complexes required for nonhistone protein acetylation. In particular, we find a key role for the Ada2 and Ada3 subunits in regulating acetylations on our fusion substrate. In contrast, other subunits tested were largely dispensable, including those required for SAGA stabili...Continue Reading

References

Jun 25, 1998·Molecular and Cellular Biology·J Pérez-Martín, A D Johnson
Sep 22, 1999·Molecular and Cellular Biology·A EberharterJ L Workman
May 17, 2000·Proceedings of the National Academy of Sciences of the United States of America·J LandryR Sternglanz
Jan 5, 2002·The Journal of Biological Chemistry·Ramakrishnan BalasubramanianSong Tan
Jan 5, 2002·The Journal of Biological Chemistry·David E SternerShelley L Berger
Jul 9, 2002·Molecular and Cellular Biology·Pei-Yun Jenny Wu, Fred Winston
Aug 21, 2002·Proceedings of the National Academy of Sciences of the United States of America·David E SternerShelley L Berger
Nov 26, 2002·Molecular and Cellular Biology·Marilyn G Pray-GrantPatrick A Grant
Jul 30, 2004·Molecular and Cellular Biology·Jason C TannyDanesh Moazed
Oct 21, 2004·Molecular Cell·François RobertRichard A Young
Jan 20, 2005·Molecular and Cellular Biology·Kenneth K LeeJerry L Workman
Jul 5, 2006·Molecular Cell·Nancy L MaasDavid P Toczyski
Aug 5, 2006·The EMBO Journal·Decha Sermwittayawong, Song Tan
Nov 18, 2006·EMBO Reports·Jeanne M WilsonLorraine Pillus
Jan 24, 2007·Molecular and Cellular Biology·Janet MeadAndrew K Vershon
Apr 5, 2007·Nature Protocols·Andrej ShevchenkoMatthias Mann
May 22, 2008·Genetics·Weimin PengSiavash K Kurdistani
Sep 24, 2008·Molecular and Cellular Biology·Jennifer K ChoiLeann J Howe
Jan 10, 2009·Nature Protocols·Da Wei HuangRichard A Lempicki
Feb 17, 2009·The Journal of Biological Chemistry·Shanshan Li, Michael A Shogren-Knaak
Jun 22, 2011·Molecular and Cellular Biology·Cara A Froyd, Laura N Rusche
Nov 24, 2011·Nucleic Acids Research·J Michael CherryEdith D Wong
Mar 2, 2012·Molecular Microbiology·Sandra BloemendalUlrich Kück
Mar 20, 2012·Genes & Development·Gianpiero SpedaleW W M Pim Pijnappel
Aug 7, 2012·Molecular & Cellular Proteomics : MCP·Peter HenriksenChunaram Choudhary
Apr 30, 2013·The Journal of Biological Chemistry·Tammy Pracheil, Zhengchang Liu

❮ Previous
Next ❯

Citations

Feb 26, 2020·Cancer Research·Lisa Maria MustachioSharon Y R Dent
Jul 28, 2020·Biochimica Et Biophysica Acta. Gene Regulatory Mechanisms·Michael Downey

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.

Related Papers

Current Opinion in Genetics & Development
Yannick Doyon, Jacques Côté
Proceedings of the National Academy of Sciences of the United States of America
Amrita BasuSandra B Hake
The Journal of Biological Chemistry
Ramakrishnan BalasubramanianSong Tan
© 2022 Meta ULC. All rights reserved