• Gallic acid
• Hypoparathyroidism
• Ischemia–reperfusion injury
• Mechanism

• Signal Transduction/drug effects
• Reperfusion Injury/drug therapy
• Reperfusion Injury/metabolism
• Reperfusion Injury/prevention & control
• Reperfusion Injury/pathology
• Gallic Acid/pharmacology
• Gallic Acid/analogs & derivatives
• Animals
• Apoptosis/drug effects
• Parathyroid Glands/metabolism
• Parathyroid Glands/drug effects
• Parathyroid Glands/pathology
• Cell Proliferation/drug effects
• Humans
• Mice

• 202301AY070001-009 Yunnan Provincial Science and Technology Department Science and Technology Project
• 202001AY070001-212 Yunnan Provincial Science and Technology Department Science and Technology Project
• 2023Y0654 Yunnan Provincial Education Department Scientific Research Fund
• 2022Y219 Scientific Research Foundation of Education Department of Yunnan

• Actin cytoskeleton
• Autoinflammatory disorder
• Cell cycle
• Metabolic reprogramming
• Prenylation
• Small GTPases

• Mevalonate Kinase Deficiency/genetics
• Mevalonate Kinase Deficiency/metabolism
• Mevalonate Kinase Deficiency/pathology
• Humans
• Fibroblasts/metabolism
• Fibroblasts/pathology
• Phosphotransferases (Alcohol Group Acceptor)/genetics
• Phosphotransferases (Alcohol Group Acceptor)/metabolism
• Phosphotransferases (Alcohol Group Acceptor)/deficiency
• Cells, Cultured
• Signal Transduction

• NADPH oxidase
• Rac1
• Rho5
• apoptosis
• glucose signaling
• mitochondria
• molecular switch
• oxidative stress

• Male
• Humans
• Saccharomyces cerevisiae/metabolism
• Monomeric GTP-Binding Proteins/metabolism
• Oxidative Stress
• rho GTP-Binding Proteins/metabolism
• Reactive Oxygen Species/metabolism

• POTEE
• Rac1
• SUMOylation
• TRIO-GEF
• breast cancer
• invadopodium

• Humans
• Female
• Signal Transduction
• Podosomes/metabolism
• Breast Neoplasms
• rac1 GTP-Binding Protein/metabolism
• Cell Movement
• Cell Line, Tumor

• 20410 Cancer Research UK
• A27412 Cancer Research UK
• A20971 Cancer Research UK
• Cancer Research UK
• Ministerio de Economía y Competitividad

• Dock2
• Prex1
• Rac
• Rac-GEF
• Tiam1
• Vav
• neutrophils
• small GTPase

• Humans
• Neutrophils/metabolism
• Integrins/metabolism
• rac GTP-Binding Proteins/metabolism
• 14-3-3 Proteins/metabolism
• CD18 Antigens/metabolism

• 20410 Cancer Research UK

• RAS inhibitor
• allelic RAS mutation
• cancer
• personalized therapy

• Beijing Municipal Natural Science Foundation
• National Natural Science Foundation of China

The dissociation of tissue and cell aggregates into single cells is of high interest for single cell analysis studies, primary cultures, tissue engineering, and regenerative medicine. However, current methods are slow, poorly controlled, variable, and can introduce artifacts. We previously developed a microfluidic device that contains two separate dissociation modules, a branching channel array and nylon mesh filters, which was used as a polishing step after tissue processing with a microfluidic digestion device. Here, we employed the integrated disaggregation and filtration (IDF) device as a standalone method with both cell aggregates and traditionally digested tissue to perform a well-controlled and detailed study into the effect of mechanical forces on dissociation, including modulation of flow rate, device pass number, and even the mechanism. Using a strongly cohesive cell aggregate model, we found that single cell recovery was highest using flow rates exceeding 40 ml/min and multiple passes through the filter module, either with or without the channel module. For minced and digested kidney tissue, recovery of diverse cell types was maximal using multiple passes through the channel module and only a single pass through the filter module. Notably, we found that epithelial cell recovery from the optimized IDF device alone exceeded our previous efforts, and this result was maintained after reducing digestion time to 20 min. However, endothelial cells and leukocytes still required extended digestion time for maximal recover. These findings highlight the significance of parameter optimization to achieve the highest cell yield and viability based on tissue sample size, extracellular matrix content, and strength of cell-cell interactions.

• dissociation
• microfluidics
• single cell analysis
• tissue engineering
• tissue processing

Rho family small GTPases (Rho) regulate various cell motility processes by spatiotemporally controlling the actin cytoskeleton. Some Rho-specific guanine nucleotide exchange factors (RhoGEFs) are regulated via tyrosine phosphorylation by Src family tyrosine kinase (SFK). We also previously reported that PLEKHG2, a RhoGEF for the GTPases Rac1 and Cdc42, is tyrosine-phosphorylated by SRC. However, the details of the mechanisms by which SFK regulates RhoGEFs are not well understood. In this study, we found for the first time that PLEKHG1, which has very high homology to the Dbl and pleckstrin homology domains of PLEKHG2, activates Cdc42 following activation by FYN, a member of the SFK family. We also show that this activation of PLEKHG1 by FYN requires interaction between these two proteins and FYN-induced tyrosine phosphorylation of PLEKHG1. We also found that the region containing the Src homology 3 and Src homology 2 domains of FYN is required for this interaction. Finally, we demonstrated that tyrosine phosphorylation of Tyr-720 and Tyr-801 in PLEKHG1 is important for the activation of PLEKHG1. These results suggest that FYN is a regulator of PLEKHG1 and may regulate cell morphology through Rho signaling via the interaction with and tyrosine phosphorylation of PLEKHG1.

• FYN
• Rho (Rho GTPase)
• Src family tyrosine kinase (SFK)
• Src homology 2 domain (SH2)
• cell signaling
• guanine nucleotide exchange factor (GEF)
• phosphorylation
• protein-protein interaction
• serum response factor (SRF)

• Phosphorylation
• Rho Guanine Nucleotide Exchange Factors/genetics
• Rho Guanine Nucleotide Exchange Factors/metabolism
• Tyrosine/metabolism
• rho GTP-Binding Proteins/genetics
• rho GTP-Binding Proteins/metabolism
• src-Family Kinases/genetics
• src-Family Kinases/metabolism

• Immunity
• NLR
• OsRac1
• PRR
• rice

• GTP Phosphohydrolases/genetics
• GTP Phosphohydrolases/metabolism
• NLR Proteins/genetics
• NLR Proteins/metabolism
• Oryza/genetics
• Oryza/metabolism
• Plant Immunity/genetics
• Plant Proteins/genetics
• Plant Proteins/metabolism
• Receptors, Pattern Recognition/genetics
• Receptors, Pattern Recognition/metabolism

• Hepatocellular carcinoma
• Invadopodia formation
• PI3K/Akt
• PIP2
• T-lymphoma invasion and metastasis 1

• PI3-Kinase
• Ras oncogenes

• Cell Movement/genetics
• Cell Proliferation/genetics
• Cell Transformation, Neoplastic/genetics
• Genes, ras/genetics
• Humans
• Mutation
• Neoplasms/genetics
• Neoplasms/metabolism
• Phosphatidylinositol 3-Kinases/genetics
• Phosphatidylinositol 3-Kinases/metabolism
• Signal Transduction/genetics
• ras Proteins/genetics
• ras Proteins/metabolism

• Alkyl-ether-lipid
• Breast cancer cell migration
• Constitutive Ca(2+) entry
• TRPV2

• Amino Acid Sequence
• Cell Membrane/metabolism
• GTP-Binding Proteins/metabolism
• Guanine Nucleotide Dissociation Inhibitors/chemistry
• HEK293 Cells
• Humans
• Monomeric GTP-Binding Proteins/metabolism
• Protein Binding
• rho GTP-Binding Proteins/chemistry
• rho Guanine Nucleotide Dissociation Inhibitor alpha/metabolism
• rho Guanine Nucleotide Dissociation Inhibitor alpha/physiology
• rho Guanine Nucleotide Dissociation Inhibitor beta/metabolism
• rho Guanine Nucleotide Dissociation Inhibitor beta/physiology
• rho Guanine Nucleotide Dissociation Inhibitor gamma/metabolism
• rho Guanine Nucleotide Dissociation Inhibitor gamma/physiology
• rho-Specific Guanine Nucleotide Dissociation Inhibitors/metabolism
• rho-Specific Guanine Nucleotide Dissociation Inhibitors/physiology

• G0700057 Medical Research Council
• MR/J007803/1 Medical Research Council
• BB/JO1454/1 Biotechnology and Biological Sciences Research Council
• Isaac Newton Trust
• Ministry of Higher Education, Malaysia

• adhesive crosstalk
• biomaterials
• cadherins
• integrins
• mechanosensing

Calmodulin is a ubiquitous signalling protein that controls many biological processes due to its capacity to interact and/or regulate a large number of cellular proteins and pathways, mostly in a Ca²⁺-dependent manner. This complex interactome of calmodulin can have pleiotropic molecular consequences, which over the years has made it often difficult to clearly define the contribution of calmodulin in the signal output of specific pathways and overall biological response. Most relevant for this review, the ability of calmodulin to influence the spatiotemporal signalling of several small GTPases, in particular KRas and Rac1, can modulate fundamental biological outcomes such as proliferation and migration. First, direct interaction of calmodulin with these GTPases can alter their subcellular localization and activation state, induce post-translational modifications as well as their ability to interact with effectors. Second, through interaction with a set of calmodulin binding proteins (CaMBPs), calmodulin can control the capacity of several guanine nucleotide exchange factors (GEFs) to promote the switch of inactive KRas and Rac1 to an active conformation. Moreover, Rac1 is also an effector of KRas and both proteins are interconnected as highlighted by the requirement for Rac1 activation in KRas-driven tumourigenesis. In this review, we attempt to summarize the multiple layers how calmodulin can regulate KRas and Rac1 GTPases in a variety of cellular events, with biological consequences and potential for therapeutic opportunities in disease settings, such as cancer.

• Calmodulin
• KRas
• Rac1
• calmodulin-binding proteins
• signalling

• Antineoplastic Agents/pharmacology
• Apoptosis/drug effects
• Caspases/genetics
• Caspases/metabolism
• Cell Line, Tumor
• Cell Proliferation/drug effects
• Cell Survival/drug effects
• GTP Phosphohydrolases/genetics
• GTP Phosphohydrolases/metabolism
• Gene Expression Regulation, Enzymologic/drug effects
• Gene Expression Regulation, Neoplastic/drug effects
• Humans
• Quinacrine/pharmacology

• Alzheimer’s disease
• Rac1
• forgetting
• hippocampus
• memory loss

• Animals
• Axons/metabolism
• Cell Movement/genetics
• Cells, Cultured
• Exocytosis/genetics
• GTP Phosphohydrolases/genetics
• Humans
• Mice
• Mice, Transgenic
• Peripheral Nervous System/growth & development
• Peripheral Nervous System/metabolism
• Schwann Cells/metabolism
• Signal Transduction/genetics
• ral GTP-Binding Proteins/genetics

• Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung

Binding of angiopoietin-1 (Ang-1) to its receptor Tie2 on endothelial cells (ECs) promotes vessel barrier integrity and angiogenesis. Here, we identify PAK2 and paxillin as critical targets of Ang-1 responsible for EC migration, polarization, and sprouting. We found that Ang-1 increases PAK2-dependent paxillin phosphorylation and remodeling of focal adhesions and that PAK2 and paxillin are required for EC polarization, migration, and angiogenic sprouting in response to Ang-1. Our findings show that Ang-1 triggers Cdc42 activation at the leading edges of migrating ECs, which is dependent on PAK2 and paxillin expression. We also established that the polarity protein Par3 interacts with Cdc42 in response to Ang-1 in a PAK2- and paxillin-dependent manner. Par3 is recruited at the leading edges of migrating cells and in focal adhesion, where it forms a signaling complex with PAK2 and paxillin in response to Ang-1. These results show that Ang-1 triggers EC polarization and angiogenic sprouting through PAK2-dependent paxillin activation and remodeling of focal adhesions, which are necessary for local activation of Cdc42 and the associated polarity complex. We have shown that PAK2 controls a signaling pathway important for angiogenic sprouting that links focal adhesions to polarity signaling in ECs.

• Adherens junctions
• Cadherin switching
• Collagen
• Discoidin domain receptor
• Epithelial-mesenchymal transition
• Integrin

• ATM
• Migration
• Rac1
• Reactive oxygen species

• Animals
• Ataxia Telangiectasia Mutated Proteins/metabolism
• Cell Movement
• Enzyme Activation
• HeLa Cells
• Humans
• Oxidative Stress
• Rats
• Reactive Oxygen Species/metabolism
• rac1 GTP-Binding Protein/metabolism

• Xenopus
• cell migration
• endoderm
• gastrula
• guidance cues
• mesoderm
• motility
• tissue separation

• Human lung epithelial cells
• Physicochemical properties
• Single walled carbon nanotubes (SWCNTs)
• Synergistic effects
• User-tailoring

• P20 GM103434 NIGMS NIH HHS
• P30 GM103488 NIGMS NIH HHS
• R01 ES022968 NIEHS NIH HHS
• S10 OD016165 NIH HHS

• Cell Movement/genetics
• Cell Proliferation/genetics
• Endocytosis/genetics
• Endosomes/genetics
• Humans
• Protein Transport
• Signal Transduction/genetics
• rac1 GTP-Binding Protein/genetics
• ras Proteins/genetics

• Dendritic tree complexity
• Fluorescence resonance energy transfer (FRET)
• Hippocampal cell culture
• Neuron
• Rac1 activation assay
• Ratiometric imaging
• Rho GTPases
• Sholl analysis

• Animals
• Dendrites/enzymology
• Gene Silencing
• Hippocampus/cytology
• Hippocampus/enzymology
• Nerve Net/cytology
• Nerve Net/enzymology
• RNA, Small Interfering/biosynthesis
• RNA, Small Interfering/genetics
• Rats
• Rats, Wistar
• Transfection/methods
• rac1 GTP-Binding Protein/antagonists & inhibitors
• rac1 GTP-Binding Protein/biosynthesis
• rac1 GTP-Binding Protein/genetics

• Bioimaging
• FRET sensor
• Plant immunity
• Small GTPase

• National Natural Science Foundation in China
• Grant-in-Aid for Scientific Research (C)
• 100 Talents Plan in Chinese Academy of Sciences Program A

Barely 10-20% of patients with metastatic colorectal cancer (mCRC) receive a clinical benefit from the use of anti-EGFR monoclonal antibodies (mAbs). We hypothesized that this could depends on their efficiency to reduce Store Operated Calcium Entry (SOCE) that are known to enhance cancer cells.

In the present study, we demonstrate that SOCE promotes migration of colon cancer cell following the formation of a lipid raft ion channel complex composed of TRPC1/Orai1 and SK3 channels. Formation of this complex is stimulated by the phosphorylation of the reticular protein STIM1 by EGF and activation of the Akt pathway. Our data show that, in a positive feedback loop SOCE activates both Akt pathway and SK3 channel activity which lead to SOCE amplification. This amplification occurs through the activation of Rac1/Calpain mediated by Akt. We also show that Anti-EGFR mAbs can modulate SOCE and cancer cell migration through the Akt pathway. Interestingly, the alkyl-lipid Ohmline, which we previously showed to be an inhibitor of SK3 channel, can dissociated the lipid raft ion channel complex through decreased phosphorylation of Akt and modulation of mAbs action.

This study demonstrates that the inhibition of the SOCE-dependent colon cancer cell migration trough SK3/TRPC1/Orai1 channel complex by the alkyl-lipid Ohmline may be a novel strategy to modulate Anti-EGFR mAb action in mCRC.

• Akt signaling
• Ohmline
• SOCE
• anti-EGFR mAbs
• lipid-raft channel complex

• GTPase activating proteins (GAPs)
• Rac1
• Rac1 targeting
• cancer
• cardiovascular diseases
• guanine nucleotide dissociation inhibitors (GDIs)
• guanine nucleotide exchange factors (GEFs)
• inflammatory disorders
• kidney diseases
• neurodegenerative disorders
• post-translational modifications

• Animals
• Enzyme Activation/drug effects
• Humans
• Molecular Targeted Therapy/methods
• Nucleotides/metabolism
• Signal Transduction/drug effects
• rac1 GTP-Binding Protein/metabolism

• 20410 Cancer Research UK
• MR/L007495/1 Medical Research Council
• C5759/A12328 Cancer Research UK

p27^Kip1 (p27) is a cyclin-CDK inhibitor and negative regulator of cell proliferation. p27 also controls other cellular processes including migration and cytoplasmic p27 can act as an oncogene. Furthermore, cytoplasmic p27 promotes invasion and metastasis, in part by promoting epithelial to mesenchymal transition. Herein, we find that p27 promotes cell invasion by binding to and regulating the activity of Cortactin, a critical regulator of invadopodia formation. p27 localizes to invadopodia and limits their number and activity. p27 promotes the interaction of Cortactin with PAK1. In turn, PAK1 promotes invadopodia turnover by phosphorylating Cortactin, and expression of Cortactin mutants for PAK-targeted sites abolishes p27’s effect on invadopodia dynamics. Thus, in absence of p27, cells exhibit increased invadopodia stability due to impaired PAK1-Cortactin interaction, but their invasive capacity is reduced compared to wild-type cells. Overall, we find that p27 directly promotes cell invasion by facilitating invadopodia turnover via the Rac1/PAK1/Cortactin pathway.

DOI:http://dx.doi.org/10.7554/eLife.22207.001

When animals develop from embryos to adults, or try to heal wounds later in life, their cells have to move. Moving means that the cells must invade into their surroundings, a dense network of proteins called the extracellular matrix. The cell first attaches to the extracellular matrix; degrades it; and then moves into the newly opened space. Cells have developed specialized structures called invadosomes to enable all these steps. Invadosomes are never static, they first assemble where cells interact with extracellular matrix, they then release proteins that loosen the matrix, and finally disassemble again to allow cells to move. Invadosomes in cancer cells often become overactive, and can allow the tumor cells to spread throughout the body.

A lot of different proteins are involved in controlling how and when cells move. p27 is a well-known protein usually found in a cell’s nucleus along with the cell’s DNA. Inside the nucleus, p27 suppresses tumor growth by stopping cells from dividing. However, often in cancer cells p27 moves outside of the cell’s nucleus where it contributes to cell movement via an unknown mechanism.

To answer how p27 controls cell invasion, Jeannot et al. used a biochemical technique to uncover which proteins p27 binds to when it is outside of the nucleus. One of its interaction partners was called Cortactin. This protein is known to be an important building block of invadosomes, and is involved in both the assembly and disassembly of these structures. In further experiments, Jeannot studied mouse cells with or without p27 and human cancer cells that can be grown in the laboratory. The experiments revealed that p27 promotes an enzyme called PAK1 to also bind to Cortactin. PAK1 then modified Cortactin, causing whole invadosomes to disassemble, which in turn allowed cells to de-attach from the matrix and move forward. In contrast, cells lacking p27 had more stable invadosomes, attached more strongly to the matrix and were better at degrading it, but could not invade as well as cells with p27.

Overall these experiments showed a new way that p27 promotes cell invasion. The next steps will include finding out exactly how the modification of Cortactin causes the invadosomes to disassemble. Furthermore, it will be important to study whether forcing p27 back into the nucleus can reduce the spread of cancer cells in the body.

DOI:http://dx.doi.org/10.7554/eLife.22207.002

• PAK
• cancer biology
• cell biology
• cortactin
• human
• invadopodia
• invasion
• mouse
• p27/Kip1

• TGFbeta signaling
• cancer biology
• cancer stem cells
• developmental biology
• mouse
• squamous cell carcinoma
• stem cells
• transition zones

• Adaptor Proteins, Signal Transducing/metabolism
• Animals
• Carcinoma, Squamous Cell/physiopathology
• Cell Movement
• Disease Models, Animal
• Gene Expression Regulation, Neoplastic
• Mice
• Neoplasm Metastasis
• Neoplastic Stem Cells/physiology
• Protein Serine-Threonine Kinases/deficiency
• Receptor, Transforming Growth Factor-beta Type II
• Receptors, Transforming Growth Factor beta/deficiency

• P30 AR047363 NIAMS NIH HHS
• P30 DK078392 NIDDK NIH HHS
• P30 DK090971 NIDDK NIH HHS
• Fondation ARC pour la Recherche sur le Cancer
• V Foundation for Cancer Research
• Sidney Kimmel Foundation for Cancer Research

Two pharmacologically distinct types of local protein synthesis are required for synapse- specific long-term synaptic facilitation (LTF) in Aplysia: one for initiation and the other for maintenance. ApCPEB, a rapamycin sensitive prion-like molecule regulates a form of local protein synthesis that is specifically required for the maintenance of the LTF. However, the molecular component of the local protein synthesis that is required for the initiation of LTF and that is sensitive to emetine is not known. Here, we identify a homolog of ApCPEB responsible for the initiation of LTF. ApCPEB4 which we have named after its mammalian CPEB4-like homolog lacks a prion-like domain, is responsive to 5-hydroxytryptamine, and is translated (but not transcribed) in an emetine-sensitive, rapamycin-insensitive, and PKA-dependent manner. The ApCPEB4 binds to different target RNAs than does ApCPEB. Knock-down of ApCPEB4 blocked the induction of LTF, whereas overexpression of ApCPEB4 reduces the threshold of the formation of LTF. Thus, our findings suggest that the two different forms of CPEBs play distinct roles in LTF; ApCPEB is required for maintenance of LTF, whereas the ApCPEB4, which lacks a prion-like domain, is required for the initiation of LTF.

• Aplysia
• CPEB
• CPEB4
• Long-term facilitation

• Tiam1
• immunohistochemistry
• ovarian carcinoma
• prognosis
• survival analysis

• Guanine nucleotide Exchange Factors (GEFs)
• IQGAP1
• Ingenuity Integrated Pathway Analysis (IPA)
• P-Rex1
• Rac1
• Small GTPases
• Stable Isotope Labeling by Amino acids in Cell culture (SILAC)
• TMOD3
• Tandem Affinity Purification (TAP)
• Tiam1

• Animals
• Cell Communication
• Cell Line, Tumor
• Cell Movement
• Guanine Nucleotide Exchange Factors/metabolism
• Humans
• Isotope Labeling
• Mice
• NIH 3T3 Cells
• Protein Binding
• Protein Interaction Mapping
• Proteomics/methods
• Signal Transduction
• rac1 GTP-Binding Protein/metabolism
• ras GTPase-Activating Proteins/metabolism

• 20410 Cancer Research UK

• MDCK cells
• epithelial-to-mesenchymal transition
• fiber diameter
• fibrous scaffolds
• hepatocyte growth factor
• phenotype

• Animals
• Biomimetic Materials/chemistry
• Cell Adhesion/physiology
• Cell Adhesion Molecules/physiology
• Cell Growth Processes/drug effects
• Cell Growth Processes/physiology
• Cellular Microenvironment/physiology
• Dogs
• Epithelial Cells/cytology
• Epithelial Cells/drug effects
• Epithelial-Mesenchymal Transition/drug effects
• Epithelial-Mesenchymal Transition/physiology
• Hepatocyte Growth Factor/pharmacology
• Madin Darby Canine Kidney Cells
• Polyesters/chemistry
• Tissue Scaffolds

• P20 GM103446 NIGMS NIH HHS
• P30 GM110758 NIGMS NIH HHS
• R01 DC014461 NIDCD NIH HHS
• R01 DE022969 NIDCR NIH HHS

Aurora A is a serine/threonine kinase that contributes to the progression of mitosis by inducing microtubule nucleation. Here we have identified an unexpected role for Aurora A kinase in antigen-driven T-cell activation. We find that Aurora A is phosphorylated at the immunological synapse (IS) during TCR-driven cell contact. Inhibition of Aurora A with pharmacological agents or genetic deletion in human or mouse T cells severely disrupts the dynamics of microtubules and CD3ζ-bearing vesicles at the IS. The absence of Aurora A activity also impairs the activation of early signalling molecules downstream of the TCR and the expression of IL-2, CD25 and CD69. Aurora A inhibition causes delocalized clustering of Lck at the IS and decreases phosphorylation levels of tyrosine kinase Lck, thus indicating Aurora A is required for maintaining Lck active. These findings implicate Aurora A in the propagation of the TCR activation signal.

Aurora A is a protein kinase that contributes to the progression of mitosis by stimulating microtubule nucleation. Here the authors show that Aurora A also functions during T cell activation by maintaining TCR signaling through Lck activation.

• E-cadherin
• adhesion
• beta-catenin
• siRNA
• transfection