scholarly journals Fluctuations in AKT and PTEN Activity Are Linked by the E3 Ubiquitin Ligase cCBL

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2803
Author(s):  
Manuel Olazábal-Morán ◽  
Miriam Sánchez-Ortega ◽  
Laura Martínez-Muñoz ◽  
Carmen Hernández ◽  
Manuel S. Rodríguez ◽  
...  
Keyword(s):  
Growth Factors ◽  
Interaction Analysis ◽  
Pi3 Kinase ◽  
Dependent Manner ◽  
Protein Protein Interaction ◽  
Tensin Homolog ◽  
Ubiquitin E3 Ligase ◽  
B Lineage ◽  
And Migration ◽  
Phosphoinositide 3 Kinase

3-poly-phosphoinositides (PIP3) regulate cell survival, division, and migration. Both PI3-kinase (phosphoinositide-3-kinase) and PTEN (phosphatase and tensin-homolog in chromosome 10) control PIP3 levels, but the mechanisms connecting PI3-kinase and PTEN are unknown. Using non-transformed cells, the activation kinetics of PTEN and of the PIP3-effector AKT were examined after the addition of growth factors. Both epidermal growth factor and serum induced the early activation of AKT and the simultaneous inactivation of PTEN (at ~5 min). This PIP3/AKT peak was followed by a general reduction in AKT activity coincident with the recovery of PTEN phosphatase activity (at ~10–15 min). Subsequent AKT peaks and troughs followed. The fluctuation in AKT activity was linked to that of PTEN; PTEN reconstitution in PTEN-null cells restored AKT fluctuations, while PTEN depletion in control cells abrogated them. The analysis of PTEN activity fluctuations after the addition of growth factors showed its inactivation at ~5 min to be simultaneous with its transient ubiquitination, which was regulated by the ubiquitin E3 ligase cCBL (casitas B-lineage lymphoma proto-oncogene). Protein-protein interaction analysis revealed cCBL to be brought into the proximity of PTEN in a PI3-kinase-dependent manner. These results reveal a mechanism for PI3-kinase/PTEN crosstalk and suggest that cCBL could be new target in strategies designed to modulate PTEN activity in cancer.

scholarly journals Rac1-mediated membrane raft localization of PI3K/p110β is required for its activation by GPCRs or PTEN loss

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Onur Cizmecioglu ◽  
Jing Ni ◽  
Shaozhen Xie ◽  
Jean J Zhao ◽  
Thomas M Roberts
Keyword(s):  
Plasma Membrane ◽  
Physiological Role ◽  
Dependent Manner ◽  
Membrane Raft ◽  
Cell Growth And Migration ◽  
Gpcr Signaling ◽  
Pten Loss ◽  
And Migration ◽  
Phosphoinositide 3 Kinase ◽  
Genetic Targeting

We aimed to understand how spatial compartmentalization in the plasma membrane might contribute to the functions of the ubiquitous class IA phosphoinositide 3-kinase (PI3K) isoforms, p110α and p110β. We found that p110β localizes to membrane rafts in a Rac1-dependent manner. This localization potentiates Akt activation by G-protein-coupled receptors (GPCRs). Thus genetic targeting of a Rac1 binding-deficient allele of p110β to rafts alleviated the requirement for p110β-Rac1 association for GPCR signaling, cell growth and migration. In contrast, p110α, which does not play a physiological role in GPCR signaling, is found to reside in nonraft regions of the plasma membrane. Raft targeting of p110α allowed its EGFR-mediated activation by GPCRs. Notably, p110β dependent, PTEN null tumor cells critically rely upon raft-associated PI3K activity. Collectively, our findings provide a mechanistic account of how membrane raft localization regulates differential activation of distinct PI3K isoforms and offer insight into why PTEN-deficient cancers depend on p110β.

scholarly journals 8-Hydroxydaidzein Downregulates JAK/STAT, MMP, Oxidative Phosphorylation, and PI3K/AKT Pathways in K562 Cells

Biomedicines ◽  
2021 ◽  
Vol 9 (12) ◽  
pp. 1907
Author(s):  
Pei-Shan Wu ◽  
Chih-Yang Wang ◽  
Pin-Shern Chen ◽  
Jui-Hsiang Hung ◽  
Jui-Hung Yen ◽  
...  
Keyword(s):  
Oxidative Phosphorylation ◽  
Cancer Progression ◽  
Interaction Analysis ◽  
K562 Cells ◽  
Systematic Investigation ◽  
Janus Kinase ◽  
Akt Phosphorylation ◽  
Protein Protein Interaction ◽  
Survival Signaling ◽  
Phosphoinositide 3 Kinase

A metabolite isolated from fermented soybean, 8-hydroxydaidzein (8-OHD, 7,8,4′-trihydroxyisoflavone, NSC-678112), is widely used in ethnopharmacological research due to its anti-proliferative and anti-inflammatory effects. We reported previously that 8-OHD provoked reactive oxygen species (ROS) overproduction, and induced autophagy, apoptosis, breakpoint cluster region-Abelson murine leukemia viral oncogene (BCR-ABL) degradation, and differentiation in K562 human chronic myeloid leukemia (CML) cells. However, how 8-OHD regulates metabolism, the extracellular matrix during invasion and metastasis, and survival signaling pathways in CML remains largely unexplored. High-throughput technologies have been widely used to discover the therapeutic targets and pathways of drugs. Bioinformatics analysis of 8-OHD-downregulated differentially expressed genes (DEGs) revealed that Janus kinase/signal transducer and activator of transcription (JAK/STAT), matrix metalloproteinases (MMPs), c-Myc, phosphoinositide 3-kinase (PI3K)/AKT, and oxidative phosphorylation (OXPHOS) metabolic pathways were significantly altered by 8-OHD treatment. Western blot analyses validated that 8-OHD significantly downregulated cytosolic JAK2 and the expression and phosphorylation of STAT3 dose- and time-dependently in K562 cells. Zymography and transwell assays also confirmed that K562-secreted MMP9 and invasion activities were dose-dependently inhibited by 8-OHD after 24 h of treatment. RT-qPCR analyses verified that 8-OHD repressed metastasis and OXPHOS-related genes. In combination with DisGeNET, it was found that 8-OHD’s downregulation of PI3K/AKT is crucial for controlling CML development. A STRING protein–protein interaction analysis further revealed that AKT and MYC are hub proteins for cancer progression. Western blotting revealed that AKT phosphorylation and nuclear MYC expression were significantly inhibited by 8-OHD. Collectively, this systematic investigation revealed that 8-OHD exerts anti-CML effects by downregulating JAK/STAT, PI3K/AKT, MMP, and OXPHOS pathways, and MYC expression. These results could shed new light on the development of 8-OHD for CML therapy.

scholarly journals Transcriptome-IPMS analysis reveals a tissue-dependent miR156/SPL13 regulatory mechanism in alfalfa drought tolerance

2020 ◽  
Author(s):  
Biruk A Feyissa ◽  
Justin Renaud ◽  
Vida Nasrollahi ◽  
Susanne Kohalmi ◽  
Abdelali Hannoufa
Keyword(s):  
Drought Stress ◽  
Metal Ion ◽  
Interaction Analysis ◽  
Expression Patterns ◽  
Specific Gene ◽  
Primary Metabolism ◽  
Dependent Manner ◽  
Protein Protein Interaction ◽  
Electron Transport Chains ◽  
Transcript Profiles

Abstract Background We previously reported on the interplay between miR156/SPL13 and WD40-1/DFR to improve response to drought stress in alfalfa (Medicago sativa L.). Here we aimed to investigate whether the role of miR156/SPL13 module in drought response is tissue-specific, and to identify SPL13-interacting proteins. We analyzed the global transcript profiles of leaf, stem, and root tissues of one-month old RNAi-silenced SPL13 (SPL13RNAi) alfalfa plants exposed to drought stress and conducted protein-protein interaction analysis to identify SPL13 interacting partners . Result Transcript analysis combined with weighted gene co-expression network analysis showed tissue and genotype-specific gene expression patterns. Moreover, pathway analysis of stem-derived differentially expressed genes (DEG) revealed upregulation of genes associated with stress mitigating primary and specialized metabolites, whereas genes associated with photosynthesis light reactions were silenced in SPL13RNAi plants. Leaf-derived DEG were attributed to enhanced light reactions, largely photosystem I, II, and electron transport chains, while roots of SPL13RNAi plants upregulated transcripts associated with metal ion transport, carbohydrate, and primary metabolism. Using immunoprecipitation combined with mass spectrometry (IPMS) we showed that SPL13 interacts with proteins involved in photosynthesis, specialized metabolite biosynthesis, and stress tolerance. Conclusions We conclude that the miR156/SPL13 module mitigates drought stress in alfalfa by regulating molecular and physiological processes in a tissue-dependent manner.

The phosphoinositide 3-kinase pathway and cancer

2005 ◽  
Vol 7 (10) ◽  
pp. 1-22 ◽  
Author(s):  
David Stokoe
Keyword(s):  
Signal Transduction ◽  
Kinase Activity ◽  
Malignant Potential ◽  
Pi3 Kinase ◽  
Signal Transduction Pathways ◽  
Constitutive Activation ◽  
And Migration ◽  
Phosphoinositide 3 Kinase ◽  
Pi3 Kinase Pathway ◽  
Kinase Pathway

Human tumours emerge as the result of multiple genetic and epigenetic aberrations that allow the proto-cancer cell to escape normal social control. Many signal transduction pathways become constitutively active during this process, and one whose importance is increasingly being appreciated involves phosphoinositide 3-kinase (PI3-kinase). This pathway normally regulates important cell decisions such as growth, division, survival and migration, and when deregulated it can confer malignant potential to the ensuing tumour. However, constitutive activation of the PI3-kinase pathway might provide attractive therapeutic targets for the design of small-molecule inhibitors. This review discusses events upstream and downstream of PI3-kinase activity in the PI3-kinase signalling pathway, how PI3-kinase is deregulated in human tumourigenesis, and how this is being targeted clinically.

scholarly journals Protein-protein Interaction Analysis of Contributing Molecules in Dura mater Healing Process

2020 ◽  
Vol 13 (3) ◽  
pp. 73-82
Author(s):  
Hendrikus Masang Ban Bolly ◽  
Yulius Hermanto ◽  
Ahmad Faried ◽  
Muhammad Zafrullah Arifin ◽  
Trajanus Laurens Yembise ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Dura Mater ◽  
Protein Interaction ◽  
Confidence Level ◽  
Interaction Analysis ◽  
Healing Process ◽  
Brain Anatomy ◽  
Mesenchymal Transition ◽  
Protein Protein Interaction

Background: Dura mater is a special tissue that fulfills a critical function in brain anatomy and physiology. This tissue contains numerous cells, stem cells, and growth factors. This research investigates the protein interaction contributing to dura mater healing process. Methods: We use the available analysis software to perform the protein-protein interaction (PPI) analysis (http://gpsprot.org/index.php). GPS Protein is an interactive platform for visualizing human protein interaction by integrating HIPPIE and CORUM databases. We excluded HIV-1 proteomic and RNAi databases, instead focused on human PPI (Confidence level 0.75). Two proteins were inputted as query to identify the potential protein network in Dura mater healing according to previous studies, i.e. fibroblast growth factor-2 (FGF2) and transforming growth factor beta-1 (TGFβ1). Results: PPI results shows a high level (confidence level > 0.75) of protein-protein interaction of TGFβ1 to 197 other proteins (Confidence level ranges: 0.49 - 0.87), and PPI of FGF2 to 26 other proteins (Confidence level ranges: 0.0-0.97). TGFβ1 interactions showed the important interactions to some remodeling proteins. TGFβ1 encoded regulates cell proliferation, differentiation, growth, expression modulation and the activation of other growth factors. It also induces epithelial-to-mesenchymal transition (EMT) and cell migration. Conclusion: This bioinformatics approach is the more efficient and cheaper method for analyzing the molecular aspect of protein that has a special contribution in Dura mater healing process. These results could beneficial in focusing further researches for more complex laboratory examinations.

scholarly journals Transcriptome-IPMS analysis reveals a tissue-dependent miR156/SPL13 regulatory mechanism in alfalfa drought tolerance

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Biruk A. Feyissa ◽  
Justin Renaud ◽  
Vida Nasrollahi ◽  
Susanne E. Kohalmi ◽  
Abdelali Hannoufa
Keyword(s):  
Drought Stress ◽  
Metal Ion ◽  
Interaction Analysis ◽  
Expression Patterns ◽  
Specific Gene ◽  
Primary Metabolism ◽  
Dependent Manner ◽  
Protein Protein Interaction ◽  
Electron Transport Chains ◽  
Transcript Profiles

Abstract Background We previously reported on the interplay between miR156/SPL13 and WD40–1/DFR to improve response to drought stress in alfalfa (Medicago sativa L.). Here we aimed to investigate whether the role of miR156/SPL13 module in drought response is tissue-specific, and to identify SPL13-interacting proteins. We analyzed the global transcript profiles of leaf, stem, and root tissues of one-month old RNAi-silenced SPL13 (SPL13RNAi) alfalfa plants exposed to drought stress and conducted protein-protein interaction analysis to identify SPL13 interacting partners. Result Transcript analysis combined with weighted gene co-expression network analysis showed tissue and genotype-specific gene expression patterns. Moreover, pathway analysis of stem-derived differentially expressed genes (DEG) revealed upregulation of genes associated with stress mitigating primary and specialized metabolites, whereas genes associated with photosynthesis light reactions were silenced in SPL13RNAi plants. Leaf-derived DEG were attributed to enhanced light reactions, largely photosystem I, II, and electron transport chains, while roots of SPL13RNAi plants upregulated transcripts associated with metal ion transport, carbohydrate, and primary metabolism. Using immunoprecipitation combined with mass spectrometry (IPMS) we showed that SPL13 interacts with proteins involved in photosynthesis, specialized metabolite biosynthesis, and stress tolerance. Conclusions We conclude that the miR156/SPL13 module mitigates drought stress in alfalfa by regulating molecular and physiological processes in a tissue-dependent manner.

scholarly journals FoxO6 transcriptional activity is regulated by Thr26 and Ser184, independent of nucleo-cytoplasmic shuttling

2005 ◽  
Vol 391 (3) ◽  
pp. 623-629 ◽  
Author(s):  
Lars P. van der Heide ◽  
Frank M. J. Jacobs ◽  
J. Peter H. Burbach ◽  
Marco F. M. Hoekman ◽  
Marten P. Smidt
Keyword(s):  
Transcription Factor ◽  
Growth Factors ◽  
Transcriptional Activity ◽  
Dependent Manner ◽  
Binding Characteristics ◽  
Forkhead Domain ◽  
Regulation Of Metabolism ◽  
Active Transcription ◽  
Phosphoinositide 3 Kinase ◽  
Constitutively Active

Forkhead members of the ‘O’ class (FoxO) are transcription factors crucial for the regulation of metabolism, cell cycle, cell death and cell survival. FoxO factors are regulated by insulin-mediated activation of PI3K (phosphoinositide 3-kinase)–PKB (protein kinase B) signalling. Activation of PI3K–PKB signalling results in the phosphorylation of FoxO factors on three conserved phosphorylation motifs, which are essential for the translocation of FoxO factors from the nucleus to the cytosol. FoxO6, however, remains mostly nuclear due to the fact that its shuttling ability is dramatically impaired. FoxO1, FoxO3 and FoxO4 all contain an N- and C-terminal PKB motif and a motif located in the forkhead domain. FoxO6 lacks the conserved C-terminal PKB motif, which is the cause of the shuttling impairment. Since FoxO6 can be considered constitutively nuclear, we investigated whether it is also a constitutively active transcription factor. Our results show that FoxO6 transcriptional activity is inhibited by growth factors, independent of shuttling, indicating that it is not constitutively active. The PKB site in the forkhead domain (Ser184) regulated the DNA binding characteristics and the N-terminal PKB site acted as a growth factor sensor. In summary, FoxO6 is not a constitutively active transcription factor and can be regulated by growth factors in a Thr26- and Ser184-dependent manner, independent of shuttling to the cytosol.

scholarly journals Transcriptome-IPMS analysis reveals a tissue-dependent miR156/SPL13 regulatory mechanism in alfalfa drought tolerance

2020 ◽  
Author(s):  
Biruk A Feyissa ◽  
Justin Renaud ◽  
Vida Nasrollahi ◽  
Susanne Kohalmi ◽  
Abdelali Hannoufa
Keyword(s):  
Drought Stress ◽  
Metal Ion ◽  
Interaction Analysis ◽  
Expression Patterns ◽  
Specific Gene ◽  
Primary Metabolism ◽  
Dependent Manner ◽  
Protein Protein Interaction ◽  
Electron Transport Chains ◽  
Transcript Profiles

Abstract Background : We previously reported on the interplay between miR156/SPL13 and WD40-1/DFR to improve response to drought stress in alfalfa ( Medicago sativa L.). Here we aimed to investigate whether the role of miR156/SPL13 module in drought response is tissue-specific, and to identify SPL13-interacting proteins. We analyzed the global transcript profiles of leaf, stem, and root tissues of one-month old RNAi-silenced SPL13 ( SPL13 RNAi) alfalfa plants exposed to drought stress and conducted protein-protein interaction analysis to identify SPL13 interacting partners. Result : Transcript analysis combined with weighted gene co-expression network analysis showed tissue and genotype-specific gene expression patterns. Moreover, pathway analysis of stem-derived differentially expressed genes (DEG) revealed upregulation of genes associated with stress mitigating primary and specialized metabolites, whereas genes associated with photosynthesis light reactions were silenced in SPL13 RNAi plants. Leaf-derived DEG were attributed to enhanced light reactions, largely photosystem I, II, and electron transport chains, while roots of SPL13 RNAi plants upregulated transcripts associated with metal ion transport, carbohydrate, and primary metabolism. Using immunoprecipitation combined with mass spectrometry (IPMS) we showed that SPL13 interacts with proteins involved in photosynthesis, specialized metabolite biosynthesis, and stress tolerance. Conclusions : We conclude that the miR156/SPL13 module mitigates drought stress in alfalfa by regulating molecular and physiological processes in a tissue-dependent manner.

Sign in / Sign up

Export Citation Format

Share Document