Circular Engineered Sortase for Interrogating Histone H3 in Chromatin
Reversible modification of the histone H3 N-terminal tail is critical in regulating the chromatin structure, gene expression, and cell states, while its dysregulation contributes to disease pathogenesis. Understanding the crosstalk between H3 tail modifications in nucleosomes constitutes a central challenge in epigenetics. Here, we describe an engineered sortase transpeptidase, cW11, that displays highly favorable properties for introducing scarless H3 tails onto nucleosomes. This approach significantly accelerates the production of both symmetrically and asymmetrically modified nucleosomes. We demonstrate the utility of asymmetrically modified nucleosomes produced in this way in dissecting the impact of multiple modifications on eraser enzyme processing and molecular recognition by a reader protein. Moreover, we show that cW11 sortase is very effective at cutting and tagging histone H3 tails from endogenous histones, facilitating multiplex "cut-and-paste" middle-down proteomics with tandem mass tags. This cut-and-paste proteomics approach permits the quantitative analysis of histone H3 modification crosstalk after treatment with different histone deacetylase inhibitors. We propose that these chemoenzymatic tail isolation and modification strategies made possible with cW11 sortase will broadly power epigenetic discovery and therapeutic development.
Authors
- Philip A. Cole (ORCID: https://orcid.org/0000-0001-6873-7824)
- Yingming Zhao (ORCID: https://orcid.org/0000-0003-1928-2151)
- Jennifer Jiang (ORCID: https://orcid.org/0000-0001-5186-5838)
- Kwangwoon Lee (ORCID: https://orcid.org/0000-0002-2021-5186)
- Xinlei Sheng (ORCID: https://orcid.org/0000-0002-5067-8915)
- Karim‐Jean Armache (ORCID: https://orcid.org/0000-0002-0890-9513)
- Congcong Lu
- Eunju Nam (ORCID: https://orcid.org/0000-0003-0801-9331)
- Sarah DuBois-Coyne (ORCID: https://orcid.org/0000-0001-5143-099X)
- Samuel D. Whedon (ORCID: https://orcid.org/0000-0002-3745-5645)
- Mingxuan Wu (ORCID: https://orcid.org/0000-0003-4721-0825)
- Louise Fairall (ORCID: https://orcid.org/0000-0001-9456-3047)
- Pablo De Ioannes (ORCID: https://orcid.org/0000-0003-1944-3912)
- John W. R. Schwabe (ORCID: https://orcid.org/0000-0003-2865-4383)
- Zhipeng A. Wang (ORCID: https://orcid.org/0000-0002-5693-7359)
- Emily Zahn
- Benjamin Aaron Garcia (ORCID: https://orcid.org/0000-0002-2306-1207)
- Adelina Andrei
- Emily Lundberg
- Maheeshi Yapa Abeywardana
Institutions
- Brigham and Women's Hospital (US)
- Harvard University (US)
- University of Leicester (GB)
- Washington University in St. Louis (US)
- University of Chicago (US)
- New York University (US)
- University of Pennsylvania (US)
Publication Details
- Journal
- Journal of the American Chemical Society
- Published
- 2024-11-25
- DOI
- https://doi.org/10.1021/jacs.4c12585
- Citations
- 12
- Primary Topic
- Biochemical and Structural Characterization
- Type
- article
- Field-Weighted Citation Impact
- 2.81
Funders
- National Science Foundation
- American Heart Association
- Leukemia and Lymphoma Society
- Brigham and Women's Hospital
- University of Miami
- University of Pennsylvania
- University of Chicago
- Charles A. King Trust
- Wellcome Trust
- Thermo Fisher Scientific
- York University
- University of Leicester
- Nankai University
- Perelman School of Medicine, University of Pennsylvania
- Leonard M. Miller School of Medicine
- NYU Grossman School of Medicine
- National Institute on Aging
- National Cancer Institute
- National Institute of General Medical Sciences
- National Institute of Diabetes and Digestive and Kidney Diseases