O-GlcNAcylation regulates tyrosine hydroxylase serine 40 phosphorylation and l-DOPA levels.

IF 5 2区 生物学 Q2 CELL BIOLOGY
Bruno da Costa Rodrigues, Miguel Clodomiro Dos Santos Lucena, Anna Carolina Rego Costa, Isadora de Araújo Oliveira, Mariana Thaumaturgo, Yolanda Paes-Colli, Danielle Beckman, Sergio T Ferreira, Fernando Garcia de Mello, Ricardo Augusto de Melo Reis, Adriane Regina Todeschini, Wagner Barbosa Dias
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引用次数: 0

Abstract

β-O-linked-N-acetylglucosamine (O-GlcNAcylation) is a post-translational modification (PTM) characterized by the covalent attachment of a single moiety of N-acetylglucosamine (GlcNAc) on serine/threonine residues in proteins. Tyrosine hydroxylase (TH), the rate-limiting step enzyme in the catecholamine synthesis pathway and responsible for the production of the dopamine precursor, l-3,4-dihydroxyphenylalanine (l-DOPA), has its activity regulated by phosphorylation. Here, we show an inverse feedback mechanism between O-GlcNAcylation and phosphorylation of TH at serine 40 (TH pSer40). First, we showed that, during PC12 cells neuritogenesis, TH O-GlcNAcylation decreases concurrently with the increase of pSer40. In addition, an increase in O-GlcNAcylation induces a decrease in TH pSer40 only in undifferentiated PC12 cells, whereas the decrease in O-GlcNAcylation leads to an increase in TH pSer40 levels in both undifferentiated and differentiated PC12 cells. We further show that this feedback culminates on the regulation of l-DOPA intracellular levels. Interestingly, it is noteworthy that decreasing O-GlcNAcylation is much more effective on TH pSer40 regulation than increasing its levels. Finally, ex vivo analysis confirmed the upregulation of TH pSer40 when O-GlcNAcylation levels are reduced in dopaminergic neurons from C57Bl/6 mice. Taken together, these findings demonstrate a dynamic control of l-DOPA production by a molecular cross talk between O-GlcNAcylation and phosphorylation at Ser40 in TH.NEW & NOTEWORTHY This study shows how β-O-linked-N-acetylglucosamine (O-GlcNAcylation) modulates tyrosine hydroxylase (TH) activity, revealing a negative feedback loop with Ser40 phosphorylation both in vitro and ex vivo, which directly influences on l-3,4-dihydroxyphenylalanine (l-DOPA) production. These findings offer insights into neurotransmitter homeostasis regulation, with implications for understanding and potentially treating disorders linked to aberrant catecholamine signaling.

o - glcn酰化调节酪氨酸羟化酶丝氨酸40磷酸化和左旋多巴水平。
o - glcn酰化是一种翻译后修饰,其特征是将GlcNAc的单个片段共价附着在蛋白质的丝氨酸/苏氨酸残基上。酪氨酸羟化酶(TH)是儿茶酚胺合成途径中的限速步骤酶,负责多巴胺前体L-DOPA的产生,其活性受磷酸化调节。在这里,我们展示了o - glcn酰化和TH at丝氨酸40 (TH pSer40)磷酸化之间的逆反馈机制。首先,我们发现在PC12细胞神经生成过程中,TH o - glcnac酰化水平随着pSer40水平的升高而降低。此外,o - glcn酰化的增加只会在未分化的PC12细胞中引起TH pSer40的降低,而o - glcn酰化的减少会导致未分化和已分化的PC12细胞中TH pSer40水平的升高。我们进一步表明,这种反馈在细胞内左旋多巴水平的调节上达到高潮。有趣的是,值得注意的是,减少o - glcn酰化对TH pSer40的调节比增加其水平更有效。最后,离体分析证实,当C57Bl/6小鼠多巴胺能神经元o - glcn酰化水平降低时,TH pSer40上调。综上所述,这些发现证明了酪氨酸羟化酶中o - glcn酰化和Ser40磷酸化之间的分子串扰对左旋多巴产生的动态控制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
9.10
自引率
1.80%
发文量
252
审稿时长
1 months
期刊介绍: The American Journal of Physiology-Cell Physiology is dedicated to innovative approaches to the study of cell and molecular physiology. Contributions that use cellular and molecular approaches to shed light on mechanisms of physiological control at higher levels of organization also appear regularly. Manuscripts dealing with the structure and function of cell membranes, contractile systems, cellular organelles, and membrane channels, transporters, and pumps are encouraged. Studies dealing with integrated regulation of cellular function, including mechanisms of signal transduction, development, gene expression, cell-to-cell interactions, and the cell physiology of pathophysiological states, are also eagerly sought. Interdisciplinary studies that apply the approaches of biochemistry, biophysics, molecular biology, morphology, and immunology to the determination of new principles in cell physiology are especially welcome.
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