Autoregulatory mechanism of enzyme activity by the nuclear localization signal of lysine-specific demethylase 1.

Autor: Senanayaka D; Department of Chemistry, Marquette University, Milwaukee, Wisconsin, USA., Zeng D; Department of Chemistry, Marquette University, Milwaukee, Wisconsin, USA., Alishiri S; Department of Chemistry, Marquette University, Milwaukee, Wisconsin, USA., Martin WJ; Center for Structural Biology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA; Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee, USA., Moore KI; Department of Chemistry, Marquette University, Milwaukee, Wisconsin, USA., Patel R; Department of Chemistry, Marquette University, Milwaukee, Wisconsin, USA., Luka Z; Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee, USA., Hirschi A; Center for Structural Biology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA; Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee, USA., Reiter NJ; Department of Chemistry, Marquette University, Milwaukee, Wisconsin, USA. Electronic address: nicholas.reiter@marquette.edu.
Jazyk: angličtina
Zdroj: The Journal of biological chemistry [J Biol Chem] 2024 Sep; Vol. 300 (9), pp. 107607. Date of Electronic Publication: 2024 Jul 30.
DOI: 10.1016/j.jbc.2024.107607
Abstrakt: 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 (IC 50 ∼ 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.
Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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