Autoregulatory Mechanism of Enzyme Activity by the Nuclear Localization Signal of Lysine-Specific Demethylase 1

Authors

Dulmi Senanayaka, Marquette University
Danyun Zeng, Marquette University
Sahar Alishiri, Marquette University
William J. Martin, Vanderbilt University
Khadijah I. Moore, Marquette University
Roshni Patel, Marquette University
Zigmund Luka, Vanderbilt University
Alexander Hirschi, Vanderbilt University
Nicholas J. Reiter, Marquette UniversityFollow

Document Type

Article

Publication Date

2024

Publisher

American Society for Biochemistry and Molecular Biology

Source Publication

Journal of Biological Chemistry

Source ISSN

0021-9258

Original Item ID

DOI: 10.1016/j.jbc.2024.107607

Abstract

The N-terminal region of the human lysine-specific demethylase 1 (LSD1) has no predicted structural elements, contains a nuclear localization signal (NLS), undergoes multiple posttranslational modifications (PTMs), and acts as a protein-protein interaction hub. This intrinsically disordered region (IDR) extends from core LSD1 structure, resides atop the catalytic active site, and is known to be dispensable for catalysis. Here, we show differential nucleosome binding between the full-length and an N terminus deleted LSD1 and identify that a conserved NLS and PTM containing element of the N terminus contains an alpha helical structure, and that this conserved element impacts demethylation. Enzyme assays reveal that LSD1’s own electropositive NLS amino acids 107 to 120 inhibit demethylation activity on a model histone 3 lysine 4 dimethyl (H3K4me2) peptide (K_i^app ∼ 3.3 μM) and histone 3 lysine 4 dimethyl nucleosome substrates (IC₅₀ ∼ 30.4 μM), likely mimicking the histone H3 tail. Further, when the identical, inhibitory NLS region contains phosphomimetic modifications, inhibition is partially relieved. Based upon these results and biophysical data, a regulatory mechanism for the LSD1-catalyzed demethylation reaction is proposed whereby NLS-mediated autoinhibition can occur through electrostatic interactions, and be partially relieved through phosphorylation that occurs proximal to the NLS. Taken together, the results highlight a dynamic and synergistic role for PTMs, intrinsically disordered regions, and structured regions near LSD1 active site and introduces the notion that phosphorylated mediated NLS regions can function to fine-tune chromatin modifying enzyme activity.

Comments

Published version. Journal of Biological Chemistry, Vol. 330, No. 9 (2024). DOI. © 2024 The Authors, published by Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology.

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Senanayaka, Dulmi; Zeng, Danyun; Alishiri, Sahar; Martin, William J.; Moore, Khadijah I.; Patel, Roshni; Luka, Zigmund; Hirschi, Alexander; and Reiter, Nicholas J., "Autoregulatory Mechanism of Enzyme Activity by the Nuclear Localization Signal of Lysine-Specific Demethylase 1" (2024). Chemistry Faculty Research and Publications. 1098.
https://epublications.marquette.edu/chem_fac/1098