Yeast Protein Kinase A Isoforms: A Means of Encoding Specificity in the Response to Diverse Stress Conditions?

Biomolecules, Год журнала: 2022, Номер 12(7), С. 958 - 958

Опубликована: Июль 8, 2022

Eukaryotic cells have developed a complex circuitry of signalling molecules which monitor changes in their intra- and extracellular environments. One the most widely studied pathways is highly conserved cyclic AMP (cAMP)/protein kinase A (PKA) pathway, major glucose sensing circuit yeast Saccharomyces cerevisiae. PKA activity regulates diverse targets yeast, positively activating processes that are associated with rapid cell growth (e.g., fermentative metabolism, ribosome biogenesis division) negatively regulating slow growth, such as respiratory carbohydrate storage entry into stationary phase. As higher eukaryotes, has evolved complexity at level catalytic subunit, cerevisiae expresses three isoforms, denoted Tpk1-3. Despite evidence for isoform differences multiple biological processes, molecular basis specificity remains poorly defined, many studies continue to assume redundancy regards PKA-mediated regulation. canonically been shown play key role fine-tuning cellular response stressors; however, recent now begun interrogate requirement individual isoforms coordinating distinct steps stress pathways. In this review, we discuss known non-redundant functions Tpk subunits evolving picture how these establish different conditions.

Язык: Английский

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ERK1/2 inhibitors act as monovalent degraders inducing ubiquitylation and proteasome-dependent turnover of ERK2, but not ERK1

Biochemical Journal, Год журнала: 2023, Номер 480(9), С. 587 - 605

Опубликована: Апрель 5, 2023

Innate or acquired resistance to small molecule BRAF MEK1/2 inhibitors (BRAFi MEKi) typically arises through mechanisms that sustain reinstate ERK1/2 activation. This has led the development of a range (ERKi) either inhibit kinase catalytic activity (catERKi) additionally prevent activating pT-E-pY dual phosphorylation by (dual-mechanism dmERKi). Here, we show eight different ERKi (both catERKi dmERKi) drive turnover ERK2, most abundant ERK isoform, with little no effect on ERK1. Thermal stability assays do not destabilise ERK2 (or ERK1) in vitro, suggesting is cellular consequence binding. observed upon treatment MEKi alone, it binding drives turnover. However, pre-treatment, which blocks and dissociation from MEK1/2, prevents cells poly-ubiquitylation proteasome-dependent pharmacological genetic inhibition Cullin-RING E3 ligases this. Our results suggest ERKi, including current clinical candidates, act as ‘kinase degraders’, driving their major target, ERK2. may be relevant suggestion kinase-independent effects therapeutic use ERKi.

Язык: Английский

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Cyclin D-CDK4 Disulfide Bond Attenuates Pulmonary Vascular Cell Proliferation

Circulation Research, Год журнала: 2023, Номер 133(12), С. 966 - 988

Опубликована: Ноя. 13, 2023

Pulmonary hypertension (PH) is a chronic vascular disease characterized, among other abnormalities, by hyperproliferative smooth muscle cells and perturbed cellular redox metabolic balance. Oxidants induce cell cycle arrest to halt proliferation; however, little known about the redox-regulated effector proteins that mediate these processes. Here, we report novel kinase-inhibitory disulfide bond in cyclin D-CDK4 (cyclin-dependent kinase 4) investigate its role proliferation PH.

Язык: Английский

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Oxidative stress sensing and response in neural stem cell fate

Free Radical Biology and Medicine, Год журнала: 2021, Номер 169, С. 74 - 83

Опубликована: Апрель 18, 2021

Язык: Английский

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RedOx regulation of LRRK2 kinase activity by active site cysteines

npj Parkinson s Disease, Год журнала: 2024, Номер 10(1)

Опубликована: Апрель 3, 2024

Abstract Mutations of the human leucine-rich repeat kinase 2 (LRRK2) have been associated with both, idiopathic and familial Parkinson’s disease (PD). Most these pathogenic mutations are located in domain (KD) or GTPase LRRK2. In this study we describe a mechanism which protein activity can be modulated by reversible oxidation reduction, involving unique pair adjacent cysteines, “CC” motif. Among all kinases, only LRRK2 contains motif (C2024 C2025) Activation Segment (AS) domain. an approach combining site-directed mutagenesis, biochemical analyses, cell-based assays, Gaussian accelerated Molecular Dynamics (GaMD) simulations could attribute role for each those cysteines. We employed reducing oxidizing agents potential clinical relevance to investigate effects on microtubule docking. find that cysteine gives distinct contribution: first cysteine, C2024, is essential activity, while C2025, contributes significantly redox sensitivity. Implementing thiolates (R-S - ) GaMD allowed us analyse how cysteines interacts its surrounding residues depending state. From our studies conclude downregulate hyperactive PD may provide promising tools therapeutic strategies.

Язык: Английский

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