80 research outputs found

    PROTEOMIC APPROACHES TO IDENTIFY UNIQUE AND SHARED SUBSTRATES AMONG KINASE FAMILY MEMBERS

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    Protein phosphorylation is a reversible post-translational modification that is a critical component of almost all signaling pathways. Kinases regulate substrate proteins through phosphorylation, and nearly all proteins are phosphorylated to some extent. Crucially, breakdown in phosphorylation signaling is an underlying factor in many diseases, including cancer. Understanding how phosphorylation signaling mediates cellular pathways is crucial for understanding cell biology and human disease. Targeted protein degradation (TPD) is a strategy to rapidly deplete a protein of interest (POI) and is applicable to any gene that is amenable to CRISPR-Cas9 editing. One TPD approach is the auxin-inducible degron (AID) system, which relies on the expression of an AID fusion protein and the F-box protein Tir1. Addition of auxin drives binding of the AID-POI and Tir1, resulting in rapid ubiquitination and degradation. Recently, we demonstrated that this approach can be used to study kinase-substrate relationships in a manner analogous to small-molecule inhibition using the kinase Plk1 as a proof-of-concept. Based on the results of this study, we applied AID-Tir1 protein degradation to interrogate kinase-substrate relationships for the Polo-like kinase (Plk), p21-activated kinase (PAK), and Aurora kinase families. Additionally, we made significant improvements to the CRISPR-Cas9 workflow and improved efficiency of AID-Tir1 cell line generation for kinases of interest. Targeted degradation of PAK1 revealed low PAK1 activity in HEK293 cells. Follow-up experiments showed that, while many phosphorylation sites are regulated by the group 1 PAKs, PAK1 does not regulate these pathways alone and likely has overlapping functions with the closely related kinase, PAK2. We applied AID-Tir1 to Aurora B and observed high correlation between Aurora B degradation and inhibition by the Aurora B inhibitor AZD1152, demonstrating that protein degradation is a selective approach to identify direct Aurora B substrates. We identified an uncharacterized truncated Aurora B isoform that is sufficient for Aurora B signaling in the absence of full-length Aurora B. Finally, we used fluorescent reporter proteins and Fluorescence Activated Cell Sorting (FACS) to greatly improve the efficiency of AID-Tir1 cell line generation for kinases of interest. These improvements make strides towards widespread implementation of targeted degradation as a tool to study kinase-substrate relationships

    19th SC@RUG 2022 proceedings 2021-2022

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    19th SC@RUG 2022 proceedings 2021-2022

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    Recent Developments on Mobile Ad-Hoc Networks and Vehicular Ad-Hoc Networks

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    This book presents collective works published in the recent Special Issue (SI) entitled "Recent Developments on Mobile Ad-Hoc Networks and Vehicular Ad-Hoc Networks”. These works expose the readership to the latest solutions and techniques for MANETs and VANETs. They cover interesting topics such as power-aware optimization solutions for MANETs, data dissemination in VANETs, adaptive multi-hop broadcast schemes for VANETs, multi-metric routing protocols for VANETs, and incentive mechanisms to encourage the distribution of information in VANETs. The book demonstrates pioneering work in these fields, investigates novel solutions and methods, and discusses future trends in these field

    19th SC@RUG 2022 proceedings 2021-2022

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    Process Modeling in Pyrometallurgical Engineering

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    The Special Issue presents almost 40 papers on recent research in modeling of pyrometallurgical systems, including physical models, first-principles models, detailed CFD and DEM models as well as statistical models or models based on machine learning. The models cover the whole production chain from raw materials processing through the reduction and conversion unit processes to ladle treatment, casting, and rolling. The papers illustrate how models can be used for shedding light on complex and inaccessible processes characterized by high temperatures and hostile environment, in order to improve process performance, product quality, or yield and to reduce the requirements of virgin raw materials and to suppress harmful emissions

    19th SC@RUG 2022 proceedings 2021-2022

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    19th SC@RUG 2022 proceedings 2021-2022

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    PPARs as Key Mediators of Metabolic and Inflammatory Regulation

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    Mounting evidence suggests a bidirectional relationship between metabolism and inflammation. Molecular crosstalk between these processes occurs at different levels with the participation of nuclear receptors, including peroxisome proliferator-activated receptors (PPARs). There are three PPAR isotypes, α, β/δ, and γ, which modulate metabolic and inflammatory pathways, making them key for the control of cellular, organ, and systemic processes. PPAR activity is governed by fatty acids and fatty acid derivatives, and by drugs used in clinics (glitazones and fibrates). The study of PPAR action, also modulated by post-translational modifications, has enabled extraordinary advances in the understanding of the multifaceted roles of these receptors in metabolism, energy homeostasis, and inflammation both in health and disease. This Special Issue of IJMS includes a broad range of basic and translational article, both original research and reviews, focused on the latest developments in the regulation of metabolic and/or inflammatory processes by PPARs in all organs and the microbiomes of different vertebrate species

    19th SC@RUG 2022 proceedings 2021-2022

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