Article
Cell Biology
Matthew D. Martens, Nivedita Seshadri, Lucas Nguyen, Donald Chapman, Elizabeth S. Henson, Bo Xiang, Landon Falk, Arielys Mendoza, Sunil Rattan, Jared T. Field, Philip Kawalec, Spencer B. Gibson, Richard Keijzer, Ayesha Saleem, Grant M. Hatch, Christine A. Doucette, Jason M. Karch, Vernon W. Dolinsky, Ian M. Dixon, Adrian R. West, Christof Rampitsch, Joseph W. Gordon
Summary: This study revealed the cardioprotective effect of misoprostol during neonatal hypoxic injury by modulating Bnip3 function; further demonstrated the importance of Bnip3 phosphorylation in regulating cardiomyocyte function and necroinflammation; and identified a potential pharmacological mechanism for preventing neonatal hypoxic injury.
CELL DEATH & DISEASE
(2021)
Article
Biochemistry & Molecular Biology
William M. Marsiglia, Arthur Chow, Zaigham M. Khan, Liu He, Arvin C. Dar
Summary: This study presents a NanoBRET-based assay to quantify the direct target engagement of MEK inhibitors on MEK1 and its complexes with ARAF, BRAF, CRAF, KSR1 and KSR2 in living cells. The study reveals the preferences of MEK inhibitors among these complexes and their binding profiles. Furthermore, the assay can also report on the effect of pathogenic mutations on MEK inhibitor binding. These methods are important for screening compounds targeting specific complexes in the RAS-MAPK cascade.
NATURE CHEMICAL BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Zuzana Trosanova, Petr Lousa, Aneta Kozelekova, Tomas Brom, Norbert Gasparik, Jan Tungli, Veronika Weisova, Erik Zupa, Gabriel Zoldak, Joze Hritz
Summary: This study describes the equilibrium between monomers, homo- and heterodimers of 14-3-3ζ isoform in the unmodified and phosphorylated form. The researchers found that phosphorylation of 14-3-3ζ at Ser58 significantly increases its homodimeric K-d value. Various experimental methods were used to efficiently monitor the dimerization of 14-3-3ζ protein and observe the effects of external factors and client protein binding.
JOURNAL OF MOLECULAR BIOLOGY
(2022)
Article
Plant Sciences
Yuqing Zhu, Wei Kuang, Jun Leng, Xue Wang, Linlin Qiu, Xiangyue Kong, Yongzhang Wang, Qiang Zhao
Summary: Apple 14-3-3 proteins play an important role in salt tolerance by modulating the expression of salt stress-related genes. MdGRF6, a member of the Md14-3-3 gene family, was found to be involved in salt response. Overexpression of MdGRF6 in transgenic tobacco and apple calli resulted in decreased salt tolerance, while RNAi-mediated silencing of MdGRF6 improved salt stress tolerance. Furthermore, the expression levels of salt stress-related genes were more strongly down-regulated in MdGRF6-overexpressing apple calli.
FRONTIERS IN PLANT SCIENCE
(2023)
Article
Biochemistry & Molecular Biology
Claire C. Munier, Leonardo De Maria, Karl Edman, Anders Gunnarsson, Marianna Longo, Carol MacKintosh, Saleha Patel, Arjan Snijder, Lisa Wissler, Luc Brunsveld, Christian Ottmann, Matthew W. D. Perry
Summary: The glucocorticoid receptor (GR) is a ligand-dependent transcription factor that has a central role in inflammation and is modulated through protein-protein interactions. Key phosphorylation sites on the GR were identified, along with the responsible kinases, MINK1 and Rho-associated protein kinase 1. The study highlights MINK1 and the GR-14-3-3 axis as potential targets for future therapeutic interventions.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2021)
Article
Chemistry, Medicinal
Ghazi Aljabal, Aik-Hong Teh, Beow Keat Yap
Summary: This study predicted several potential peptides that can stabilize the dimeric form of 14-3-3 & sigma; using in silico techniques. Peptide 3 showed the strongest binding and improved the binding of ExoS protein to 14-3-3 & sigma;. Additionally, peptide 3 slowed down the aggregation process of 14-3-3 & sigma; and increased its stability.
JOURNAL OF CHEMICAL INFORMATION AND MODELING
(2023)
Article
Multidisciplinary Sciences
Bojian Ding, Sheng Yang, Matthias Schaks, Yijun Liu, Abbigale J. Brown, Klemens Rottner, Saikat Chowdhury, Baoyu Chen
Summary: In this study, the structures of WRC, both alone and bound to Rac1, were investigated using cryogenic-electron microscopy. The findings reveal that Rac1 binds to WRC at two distinct sites, and binding to the A site activates the complex. Activation involves conformational changes that result in the release of sequestered WCA polypeptide, stimulating actin polymerization by the Arp2/3 complex. These structural insights, combined with biochemical and cellular analyses, provide a novel understanding of the regulation of the Rac1-WRC-Arp2/3-actin signaling axis in various biological processes and diseases.
NATURE COMMUNICATIONS
(2022)
Article
Chemistry, Multidisciplinary
Emira J. Visser, Priyadarshini Jaishankar, Eline Sijbesma, Marloes A. M. Pennings, Edmee M. F. Vandenboorn, Xavier Guillory, R. Jeffrey Neitz, John Morrow, Shubhankar Dutta, Adam R. Renslo, Luc Brunsveld, Michelle R. Arkin, Christian Ottmann
Summary: Small-molecule stabilization of protein-protein interactions (PPIs) is a promising strategy in chemical biology and drug discovery. In this study, a fragment-linking approach targeting the interface of 14-3-3 and a peptide derived from the estrogen receptor alpha (ERα) protein was developed. The initial hybrid molecule, supported by 20 crystal structures, was optimized to selectively stabilize the 14-3-3/ERα interaction by 25-fold. The high-resolution structures of the fragments, co-crystals, and linked fragments demonstrate a feasible strategy for developing orthosteric PPI stabilizers.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Biology
Sophie E. Ruff, Nikita Vasilyev, Evgeny Nudler, Susan K. Logan, Michael J. Garabedian
Summary: Ruff et al. demonstrate that PIM1 kinase phosphorylates androgen receptor (AR) and 14-3-3 zeta to coordinate their interaction at genes involved in cell migration and invasion, ultimately regulating the AR transcriptome in prostate cancer cells. The phosphorylation of AR and 14-3-3 zeta by PIM1 leads to their co-occupancy on chromatin and recruitment of additional co-regulatory proteins like hnRNPK and TRIM28, altering AR transcriptional activity. Overall, PIM1 phosphorylation of AR and 14-3-3 zeta is crucial for orchestrating their interaction and subsequent modulation of AR-dependent gene expression.
COMMUNICATIONS BIOLOGY
(2021)
Article
Plant Sciences
Zihao Fan, Yuqing Zhu, Wei Kuang, Jun Leng, Xue Wang, Linlin Qiu, Jiyun Nie, Yongbing Yuan, Rui-Fen Zhang, Yongzhang Wang, Qiang Zhao
Summary: This study reveals the regulatory mechanism of the MdGRF8-MdWRKY18 module in promoting salt tolerance in apple. MdGRF8 enhances the stability and transcriptional activation activity of MdWRKY18 through its interaction with MdWRKY18, thereby activating the expression of salt-related genes MdSOS2 and MdSOS3, and promoting salt tolerance in apple.
Article
Chemistry, Multidisciplinary
Phillip Zhu, Stanislau Stanisheuski, Rachel Franklin, Amber Vogel, Cat Hoang Vesely, Patrick Reardon, Nikolai N. Sluchanko, Joseph S. Beckman, P. Andrew Karplus, Ryan A. Mehl, Richard B. Cooley
Summary: 14-3-3 proteins dimerize and bind phosphorylated clients to regulate their function. By introducing stable mimics of phosphorylated amino acids, researchers can study 14-3-3 function effectively. In this study, the authors improved a previous genetic code expansion system to incorporate nonhydrolyzable phosphoserine into proteins. They successfully produced biologically relevant proteins with the modified amino acid and investigated the effects of phosphorylation on protein interactions and function.
ACS CENTRAL SCIENCE
(2023)
Article
Biochemistry & Molecular Biology
Micaela Boiero Sanders, Christopher P. Toret, Audrey Guillotin, Adrien Antkowiak, Thomas Vannier, Robert C. Robinson, Alphee Michelot
Summary: The use of different actin isoforms in eukaryotic cells and the molecular mechanisms of their segregation into distinct networks are poorly understood. By using yeast as a model, researchers found that the expression of heterologous actin causes significant reorganization of the actin cytoskeleton. However, the expression of two heterologous actin variants, each specialized in assembling a different network, can rescue cytoskeletal organization and increase resistance to external perturbation.
Article
Forestry
Miao Zhang, Ziping Yang, Dong Guo, Huiliang Li, Jiahong Zhu, Shiqing Peng, Ying Wang
Summary: In this study, 81 essential client proteins that interact with H. brasiliensis 14-3-3 proteins were identified through yeast two-hybrid screening. These proteins are involved in various processes such as plant signal transduction, metabolism, development, and NR biosynthesis. The interaction between HbGF14c and HbSRPP was confirmed in plants using bimolecular fluorescence complementation assays and in vitro with Pull-down assays. The findings provide insights into the molecular regulation mechanism of 14-3-3 proteins in NR biosynthesis and have potential applications in enhancing rubber tree production through genetic improvement.
Article
Plant Sciences
Yulia V. Mikhaylova, Roman K. Puzanskiy, Maria F. Shishova
Summary: The evolutionary history of 14-3-3 proteins in plants involved multiple duplication events, leading to the formation of at least four subfamilies. Recent duplication events and gene loss in plant lineages shaped the high diversity of 14-3-3 isoforms in plants. Coexpression data revealed functional groupings of 14-3-3 proteins in different species, suggesting a relationship between gene duplication, loss, and functional plasticity.
Review
Biochemistry & Molecular Biology
Veronika Obsilova, Tomas Obsil
Summary: Signal transduction cascades efficiently transmit chemical and/or physical signals from the extracellular environment to intracellular compartments, thereby eliciting an appropriate cellular response. 14-3-3 proteins, as a family of highly conserved scaffolding molecules, play a crucial role in modulating the function of other proteins primarily through phosphorylation-dependent mechanisms. They participate in key cellular processes such as cell-cycle control, apoptosis, signal transduction, energy metabolism, and protein trafficking. However, our understanding of the molecular mechanisms by which 14-3-3 proteins regulate their binding partners remains insufficient, despite intensive research into their protein-protein interactions. This review article aims to provide an overview of recent structural studies of 14-3-3 protein complexes in order to further explore the regulatory mechanisms of these proteins.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Biophysics
Rohit Joshi, Pavel Pohl, Dita Strachotova, Petr Herman, Tomas Obsil, Veronika Obsilova
Summary: Nedd4-2, a ubiquitin ligase, is involved in the endocytosis and degradation of target molecules through interactions with 14-3-3 proteins. The binding of 14-3-3 induces structural rearrangement of Nedd4-2, affecting the mobility and emission properties of certain domains. In contrast, the active site of the HECT domain becomes more mobile upon 14-3-3 binding.
BIOPHYSICAL JOURNAL
(2022)
Article
Biochemistry & Molecular Biology
Kristyna Bousova, Monika Zouharova, Petr Herman, Jiri Vymetal, Veronika Vetyskova, Katerina Jiraskova, Jiri Vondrasek
Summary: In this study, we found that the N-terminus of TRPM5 interacts with calcium-binding proteins CaM and S100A1, and these binding regions possess unique basic residues.
Article
Biochemistry & Molecular Biology
Raju Mandal, Klara Kohoutova, Olivia Petrvalska, Matej Horvath, Pavel Srb, Vaclav Veverka, Veronika Obsilova, Tomas Obsil
Summary: Here, we characterized the interaction between p53 and FOXO4 using NMR, chemical cross-linking, and analytical ultracentrifugation. Our findings highlight the importance of this interaction for the stability of the p53:FOXO4 complex and shed light on the intertwined functions of p53 and FOXO4 in cellular homeostasis, longevity, and stress response.
Review
Biochemistry & Molecular Biology
Veronika Obsilova, Tomas Obsil
Summary: Signal transduction cascades efficiently transmit chemical and/or physical signals from the extracellular environment to intracellular compartments, thereby eliciting an appropriate cellular response. 14-3-3 proteins, as a family of highly conserved scaffolding molecules, play a crucial role in modulating the function of other proteins primarily through phosphorylation-dependent mechanisms. They participate in key cellular processes such as cell-cycle control, apoptosis, signal transduction, energy metabolism, and protein trafficking. However, our understanding of the molecular mechanisms by which 14-3-3 proteins regulate their binding partners remains insufficient, despite intensive research into their protein-protein interactions. This review article aims to provide an overview of recent structural studies of 14-3-3 protein complexes in order to further explore the regulatory mechanisms of these proteins.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Dita Strachotova, Ales Holoubek, Katerina Wolfova, Barbora Brodska, Petr Herman
Summary: Specific C-terminal nucleophosmin (NPM) mutations are associated with acute myeloid leukaemia and cause mislocalization of NPMmut to the cytoplasm. This leads to mislocalization of NPM-interacting partners, including the tumor suppressor p53. We discovered the formation of a ternary complex consisting of NPMmut, p53, and Mdm2, with p53 acting as a bridging link between Mdm2 and NPMmut. Silencing of p53 prevented mislocalization of Mdm2 in the presence of NPMmut.
Article
Chemistry, Analytical
Dita Strachotova, Ales Holoubek, Barbora Brodska, Petr Herman
Summary: This study reports on the two-photon-induced photoconversion of EGFP, which can be permanently converted by intense IR irradiation to a form with a short fluorescence lifetime and spectrally conserved emission. The nonlinear dependence of the photoconversion efficiency on light intensity allows for accurate 3D localization of the converted volume within cellular structures, which is useful for kinetic FLIM applications.
METHODS AND APPLICATIONS IN FLUORESCENCE
(2023)
Article
Biochemistry & Molecular Biology
Olivia Petrvalska, Karolina Honzejkova, Nicola Koupilova, Petr Herman, Veronika Obsilova, Tomas Obsil
Summary: CaMKK1 and CaMKK2 are important protein kinases that play key roles in physiological and pathological processes. Complex formation affects their interaction with Ca2+/CaM and the structure of their domains, controlling their catalytic activity. The inhibitory effect of complex formation is more significant on CaMKK1 compared to CaMKK2, possibly due to the tighter and more rigid structure of the CaMKK1:14-3-3 gamma complex, which directly inhibits CaMKK1 activity.
Article
Biochemistry & Molecular Biology
Jakub Masaryk, Deepika Kale, Pavel Pohl, Francisco J. Ruiz-Castilla, Olga Zimmermannova, Veronika Obsilova, Jose Ramos, Hana Sychrova
Summary: Potassium is crucial for cellular processes, and its uptake and efflux through the plasma membrane need to be precisely regulated. The intracellular hydrophilic loop (IL2) of the Trk1 protein plays a key role in its targeting and activity regulation, and phosphorylation of specific residues within IL2 is important for these processes.
COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL
(2023)
Meeting Abstract
Biochemistry & Molecular Biology
P. Pohl, T. Obsil, V. Obsilova
Meeting Abstract
Biochemistry & Molecular Biology
K. Kohoutova, V. Docekal, A. Tekel, M. Ausserlechner, J. Vesely, T. Obsil, V. Obsilova
Meeting Abstract
Biochemistry & Molecular Biology
R. Mandal, K. Kohoutova, O. Petrvalska, M. Horvath, P. Srb, V. Veverka, V. Obsilova, T. Obsil
Meeting Abstract
Biochemistry & Molecular Biology
K. Honzejkova, V. Obsilova, T. Obsil
