Sprouty1 Prevents Cellular Senescence Maintaining Proliferation and Differentiation Capacity of Human Adipose Stem/Progenitor Cells

Abstract The role of Ras-Mitogen-activated protein kinase (MAPK) signaling in cellular aging is not precisely understood. Recently, we identified Sprouty1 (SPRY1) as a weight-loss target gene in human adipose stem/progenitor cells (ASCs) and showed that Sprouty1 is important for proper regulation of adipogenesis. In the present study, we show that loss-of-function of Sprouty1 by CRISPR/Cas9-mediated genome editing in human ASCs leads to hyper-activation of MAPK signaling and a senescence phenotype. Sprouty1 knockout ASCs undergo an irreversible cell cycle arrest, become enlarged and stain positive for senescence-associated β-galactosidase. Sprouty1 down-regulation leads to DNA double strand breaks, a considerably increased number of senescence-associated heterochromatin foci and induction of p53 and p21Cip1. In addition, we detect an increase of hypo-phosphorylated Retinoblastoma (Rb) protein in SPRY1 knockout ASCs. p16Ink4A is not induced. Moreover, we show that Sprouty1 knockout leads to induction of a senescence-associated secretory phenotype as indicated by the activation of the transcription factors NFκB and C/EBPβ and a significant increase in mRNA expression and secretion of interleukin-8 (IL-8) and CXCL1/GROα. Finally, we demonstrate that adipogenesis is abrogated in senescent SPRY1 knockout ASCs. In conclusion, this study reveals a novel mechanism showing the importance of Sprouty1 for the prevention of senescence and the maintenance of the proliferation and differentiation capacity of human ASCs.

Download Full-text

MKP-3 Has Essential Roles as a Negative Regulator of the Ras/Mitogen-Activated Protein Kinase Pathway during Drosophila Development

Molecular and Cellular Biology ◽

10.1128/mcb.24.2.573-583.2004 ◽

2004 ◽

Vol 24 (2) ◽

pp. 573-583 ◽

Cited By ~ 33

Author(s):

Myungjin Kim ◽

Guang-Ho Cha ◽

Sunhong Kim ◽

Jun Hee Lee ◽

Jeehye Park ◽

...

Keyword(s):

Protein Kinase ◽

Cell Fate ◽

Photoreceptor Cells ◽

Mapk Signaling ◽

Negative Regulator ◽

Mitogen Activated Protein Kinase ◽

Loss Of Function ◽

Wing Vein ◽

Mitogen Activated Protein

ABSTRACT Mitogen-activated protein kinase (MAPK) phosphatase 3 (MKP-3) is a well-known negative regulator in the Ras/extracellular signal-regulated kinase (ERK)-MAPK signaling pathway responsible for cell fate determination and proliferation during development. However, the physiological roles of MKP-3 and the mechanism by which MKP-3 regulates Ras/Drosophila ERK (DERK) signaling in vivo have not been determined. Here, we demonstrated that Drosophila MKP-3 (DMKP-3) is critically involved in cell differentiation, proliferation, and gene expression by suppressing the Ras/DERK pathway, specifically binding to DERK via the N-terminal ERK-binding domain of DMKP-3. Overexpression of DMKP-3 reduced the number of photoreceptor cells and inhibited wing vein differentiation. Conversely, DMKP-3 hypomorphic mutants exhibited extra photoreceptor cells and wing veins, and its null mutants showed striking phenotypes, such as embryonic lethality and severe defects in oogenesis. All of these phenotypes were highly similar to those of the gain-of-function mutants of DERK/rl. The functional interaction between DMKP-3 and the Ras/DERK pathway was further confirmed by genetic interactions between DMKP-3 loss-of-function mutants or overexpressing transgenic flies and various mutants of the Ras/DERK pathway. Collectively, these data provide the direct evidences that DMKP-3 is indispensable to the regulation of DERK signaling activity during Drosophila development.

Download Full-text

Proliferation of Peripheral Blood-derived Endothelial Progenitor Cells from Stable Angina Subjects

The Indonesian Biomedical Journal ◽

10.18585/inabj.v6i2.34 ◽

2014 ◽

Vol 6 (2) ◽

pp. 91 ◽

Cited By ~ 1

Author(s):

Yudi Her Oktaviono ◽

Djanggan Sargowo ◽

Mohammad Aris Widodo ◽

Yanni Dirgantara ◽

Angliana Chouw ◽

...

Keyword(s):

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Stable Angina ◽

Peripheral Blood ◽

Mapk Pathway ◽

Growth Factor Receptor ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Endothelial Progenitor ◽

Mitogen Activated Protein

BACKGROUND: A population of circulating Endothelial Progenitor Cells (EPCs) has been reported to play important role in maintaining endothelial function and integrity. Since EPCs culture is crucial and an optimized medium is currently available. Therefore we conducted a study to investigate whether stable angina subjects peripheral blood-derived EPCs could be cultured in this medium. Here, we performed study to detect EPCs characteristics and extracellular signalregulated kinase (Erk)1/2 Mitogen-Activated Protein Kinase (MAPK) pathway as possible underlying pathway for EPCs proliferation.METHODS: Peripheral blood EPCs from 8 stable angina subjects were cultured in an optimized medium with/without addition of supplement for 1 or 3 days. Then, the membrane of cultured EPCs were detected with immunofluorescence method for CD34, Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) and CD133. Colony forming unit (CFU) enumeration was performed. XTT Cell proliferation assay was performed to assess EPCs growth after 1 and 3-days culture. The western blot analysis was performed to detect possible activation of Erk1/2 MAPK.RESULTS: Number of EPCs and CFU cultured for 3 days were significantly higher than the ones cultured for 1 day (p=0.012). EPCs membrane markers from stable angina subjects were detected as well as CFUs were formed. There were significant increase of EPCs number, CFUs number and phosphorylated-Erk2 amount when the groups with and without supplement were compared (p<0.05). Meanwhile U0126, a MAPK Erk1/2 (MEK1/2) inhibitor, significantly inhibited the supplement-induced EPCs number, CFUs number and phosphorylated-Erk2 amount (p<0.05).CONCLUSION: Our results showed that ERK2 MAPK signaling pathway might play an important role in supplement-induced peripheral blood EPCs proliferation in subjects with stable angina.KEYWORDS: endothelial progenitor cell, EPC, p42, Erk2, proliferation

Download Full-text

Role of the Caenorhabditis elegans Shc Adaptor Protein in the c-Jun N-Terminal Kinase Signaling Pathway

Molecular and Cellular Biology ◽

10.1128/mcb.00938-08 ◽

2008 ◽

Vol 28 (23) ◽

pp. 7041-7049 ◽

Cited By ~ 19

Author(s):

Tomoaki Mizuno ◽

Kota Fujiki ◽

Aya Sasakawa ◽

Naoki Hisamoto ◽

Kunihiro Matsumoto

Keyword(s):

Caenorhabditis Elegans ◽

Signaling Pathway ◽

Adaptor Protein ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Loss Of Function ◽

Mapk Kinase ◽

Mitogen Activated Protein ◽

Npxy Motif ◽

Ptb Domain

ABSTRACT Mitogen-activated protein kinases (MAPKs) are integral to the mechanisms by which cells respond to physiological stimuli and a wide variety of environmental stresses. In Caenorhabditis elegans, the stress response is controlled by a c-Jun N-terminal kinase (JNK)-like mitogen-activated protein kinase (MAPK) signaling pathway, which is regulated by MLK-1 MAPK kinase kinase (MAPKKK), MEK-1 MAPK kinase (MAPKK), and KGB-1 JNK-like MAPK. In this study, we identify the shc-1 gene, which encodes a C. elegans homolog of Shc, as a factor that specifically interacts with MEK-1. The shc-1 loss-of-function mutation is defective in activation of KGB-1, resulting in hypersensitivity to heavy metals. A specific tyrosine residue in the NPXY motif of MLK-1 creates a docking site for SHC-1 with the phosphotyrosine binding (PTB) domain. Introduction of a mutation that perturbs binding to the PTB domain or the NPXY motif abolishes the function of SHC-1 or MLK-1, respectively, thereby abolishing the resistance to heavy metal stress. These results suggest that SHC-1 acts as a scaffold to link MAPKKK to MAPKK activation in the KGB-1 MAPK signal transduction pathway.

Download Full-text

SPRED1 Interferes with K-ras but Not H-ras Membrane Anchorage and Signaling

Molecular and Cellular Biology ◽

10.1128/mcb.00191-16 ◽

2016 ◽

Vol 36 (20) ◽

pp. 2612-2625 ◽

Cited By ~ 12

Author(s):

Elina Siljamäki ◽

Daniel Abankwa

Keyword(s):

Plasma Membrane ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Membrane Domains ◽

Loss Of Function ◽

Positive Effects ◽

Extracellular Signal ◽

Mitogen Activated Protein ◽

Legius Syndrome ◽

Novel Interactions

The Ras/mitogen-activated protein kinase (MAPK) signaling pathway is tightly controlled by negative feedback regulators, such as the tumor suppressor SPRED1. TheSPRED1gene also carries loss-of-function mutations in the RASopathy Legius syndrome. Growth factor stimulation translocates SPRED1 to the plasma membrane, triggering its inhibitory activity. However, it remains unclear whether SPRED1 there acts at the level of Ras or Raf. We show that pharmacological or galectin-1 (Gal-1)-mediated induction of B- and C-Raf-containing dimers translocates SPRED1 to the plasma membrane. This is facilitated in particular by SPRED1 interaction with B-Raf and, via its N terminus, with Gal-1. The physiological significance of these novel interactions is supported by two Legius syndrome-associated mutations that show diminished binding to both Gal-1 and B-Raf. On the plasma membrane, SPRED1 becomes enriched in acidic membrane domains to specifically perturb membrane organization and extracellular signal-regulated kinase (ERK) signaling of active K-ras4B (here, K-ras) but not H-ras. However, SPRED1 also blocks on the nanoscale the positive effects of Gal-1 on H-ras. Therefore, a combinatorial expression of SPRED1 and Gal-1 potentially regulates specific patterns of K-ras- and H-ras-dependent signaling output. More broadly, our results open up the possibility that related SPRED and Sprouty proteins act in a similar Ras and Raf isoform-specific manner.

Download Full-text

Trichoderma Mitogen-Activated Protein Kinase Signaling Is Involved in Induction of Plant Systemic Resistance

Applied and Environmental Microbiology ◽

10.1128/aem.71.10.6241-6246.2005 ◽

2005 ◽

Vol 71 (10) ◽

pp. 6241-6246 ◽

Cited By ~ 71

Author(s):

Ada Viterbo ◽

Michal Harel ◽

Benjamin A. Horwitz ◽

Ilan Chet ◽

Prasun K. Mukherjee

Keyword(s):

Protein Kinase ◽

Pseudomonas Syringae ◽

Disease Incidence ◽

Sclerotium Rolfsii ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Plant Roots ◽

Systemic Resistance ◽

Loss Of Function ◽

Mitogen Activated Protein

ABSTRACT The role of a mitogen-activated protein kinase (MAPK) TmkA in inducing systemic resistance in cucumber against the bacterial pathogen Pseudomonas syringae pv. lacrymans was investigated by using tmkA loss-of-function mutants of Trichoderma virens. In an assay where Trichoderma spores were germinated in proximity to cucumber roots, the mutants were able to colonize the plant roots as effectively as the wild-type strain but failed to induce full systemic resistance against the leaf pathogen. Interactions with the plant roots enhanced the level of tmkA transcript in T. virens and its homologue in Trichoderma asperellum. At the protein level, we could detect the activation of two forms reacting to the phospho-p44/42 MAPK antibody. Biocontrol experiments demonstrated that the tmkA mutants retain their biocontrol potential against Rhizoctonia solani in soil but are not effective against Sclerotium rolfsii in reducing disease incidence. Our results show that, unlike in many plant-pathogen interactions, Trichoderma TmkA MAPK is not involved in limited root colonization. Trichoderma, however, needs MAPK signaling in order to induce full systemic resistance in the plant.

Download Full-text

Specific Functional Features of the Cell Integrity MAP Kinase Pathway in the Dimorphic Fission Yeast Schizosaccharomyces japonicus

Journal of Fungi ◽

10.3390/jof7060482 ◽

2021 ◽

Vol 7 (6) ◽

pp. 482

Author(s):

Elisa Gómez-Gil ◽

Alejandro Franco ◽

Beatriz Vázquez-Marín ◽

Francisco Prieto-Ruiz ◽

Armando Pérez-Díaz ◽

...

Keyword(s):

Fission Yeast ◽

Environmental Changes ◽

Yeast Species ◽

Mapk Signaling ◽

Fine Tuning ◽

Mitogen Activated Protein Kinase ◽

Major Influence ◽

Cell Integrity ◽

Mitogen Activated Protein ◽

Functional Features

Mitogen activated protein kinase (MAPK) signaling pathways execute essential functions in eukaryotic organisms by transducing extracellular stimuli into adaptive cellular responses. In the fission yeast model Schizosaccharomyces pombe the cell integrity pathway (CIP) and its core effector, MAPK Pmk1, play a key role during regulation of cell integrity, cytokinesis, and ionic homeostasis. Schizosaccharomyces japonicus, another fission yeast species, shows remarkable differences with respect to S. pombe, including a robust yeast to hyphae dimorphism in response to environmental changes. We show that the CIP MAPK module architecture and its upstream regulators, PKC orthologs Pck1 and Pck2, are conserved in both fission yeast species. However, some of S. pombe’s CIP-related functions, such as cytokinetic control and response to glucose availability, are regulated differently in S. japonicus. Moreover, Pck1 and Pck2 antagonistically regulate S. japonicus hyphal differentiation through fine-tuning of Pmk1 activity. Chimeric MAPK-swapping experiments revealed that S. japonicus Pmk1 is fully functional in S. pombe, whereas S. pombe Pmk1 shows a limited ability to execute CIP functions and promote S. japonicus mycelial development. Our findings also suggest that a modified N-lobe domain secondary structure within S. japonicus Pmk1 has a major influence on the CIP signaling features of this evolutionarily diverged fission yeast.

Download Full-text

Effect of Static Magnetic Field on Monascus ruber M7 Based on Transcriptome Analysis

Journal of Fungi ◽

10.3390/jof7040256 ◽

2021 ◽

Vol 7 (4) ◽

pp. 256

Author(s):

Shuyan Yang ◽

Hongyi Zhou ◽

Weihua Dai ◽

Juan Xiong ◽

Fusheng Chen

Keyword(s):

Magnetic Field ◽

Static Magnetic Field ◽

Morphological Characteristics ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Primary Metabolism ◽

Monascus Ruber ◽

Growing Period ◽

Monascus Pigments ◽

Mitogen Activated Protein

The effects of a static magnetic field (SMF) on Monascus ruber M7 (M. ruber M7) cultured on potato dextrose agar (PDA) plates under SMF treatment at different intensities (5, 10, and 30 mT) were investigated in this paper. The results revealed that, compared with the control (CK, no SMF treatment), the SMF at all tested intensities did not significantly influence the morphological characteristics of M. ruber M7, while the intracellular and extracellular Monascus pigments (MPs) and extracellular citrinin (CIT) of M. ruber M7 were increased at 10 and 30 mT SMF but there was no impact on the MPs and CIT at 5 mT SMF. The transcriptome data of M. ruber M7 cultured at 30 mT SMF on PDA for 3 and 7 d showed that the SMF could increase the transcriptional levels of some relative genes with the primary metabolism, including the carbohydrate metabolism, amino acid metabolism, and lipid metabolism, especially in the early growing period (3 d). SMF could also affect the transcriptional levels of the related genes to the biosynthetic pathways of MPs, CIT, and ergosterol, and improve the transcription of the relative genes in the mitogen-activated protein kinase (MAPK) signaling pathway of M. ruber M7. These findings provide insights into a comprehensive understanding of the effects of SMF on filamentous fungi.

Download Full-text

The Emerging Role of Macrophages in Immune System Dysfunction under Real and Simulated Microgravity Conditions

International Journal of Molecular Sciences ◽

10.3390/ijms22052333 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2333

Author(s):

Yulong Sun ◽

Yuanyuan Kuang ◽

Zhuo Zuo

Keyword(s):

Immune System ◽

Mapk Signaling ◽

Immune Defense ◽

Mitogen Activated Protein Kinase ◽

Therapeutic Drugs ◽

Physiological Processes ◽

Immune System Dysfunction ◽

Cell Immunity ◽

Mitogen Activated Protein ◽

Microgravity Conditions

In the process of exploring space, the astronaut’s body undergoes a series of physiological changes. At the level of cellular behavior, microgravity causes significant alterations, including bone loss, muscle atrophy, and cardiovascular deconditioning. At the level of gene expression, microgravity changes the expression of cytokines in many physiological processes, such as cell immunity, proliferation, and differentiation. At the level of signaling pathways, the mitogen-activated protein kinase (MAPK) signaling pathway participates in microgravity-induced immune malfunction. However, the mechanisms of these changes have not been fully elucidated. Recent studies suggest that the malfunction of macrophages is an important breakthrough for immune disorders in microgravity. As the first line of immune defense, macrophages play an essential role in maintaining homeostasis. They activate specific immune responses and participate in large numbers of physiological activities by presenting antigen and secreting cytokines. The purpose of this review is to summarize recent advances on the dysfunction of macrophages arisen from microgravity and to discuss the mechanisms of these abnormal responses. Hopefully, our work will contribute not only to the future exploration on the immune system in space, but also to the development of preventive and therapeutic drugs against the physiological consequences of spaceflight.

Download Full-text

Topical Spilanthol Inhibits MAPK Signaling and Ameliorates Allergic Inflammation in DNCB-Induced Atopic Dermatitis in Mice

International Journal of Molecular Sciences ◽

10.3390/ijms20102490 ◽

2019 ◽

Vol 20 (10) ◽

pp. 2490 ◽

Cited By ~ 4

Author(s):

Wen-Chung Huang ◽

Chun-Hsun Huang ◽

Sindy Hu ◽

Hui-Ling Peng ◽

Shu-Ju Wu

Keyword(s):

Atopic Dermatitis ◽

Mast Cells ◽

Protein Kinase ◽

Allergic Inflammation ◽

Skin Lesions ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Mapk Pathways ◽

Mitogen Activated Protein ◽

Cox 2

Atopic dermatitis (AD) is a recurrent allergic skin disease caused by genetic and environmental factors. Patients with AD may experience immune imbalance, increased levels of mast cells, immunoglobulin (Ig) E and pro-inflammatory factors (Cyclooxygenase, COX-2 and inducible NO synthase, iNOS). While spilanthol (SP) has anti-inflammatory and analgesic activities, its effect on AD remains to be explored. To develop a new means of SP, inflammation-related symptoms of AD were alleviated, and 2,4-dinitrochlorobenzene (DNCB) was used to induce AD-like skin lesions in BALB/c mice. Histopathological analysis was used to examine mast cells and eosinophils infiltration in AD-like skin lesions. The levels of IgE, IgG1 and IgG2a were measured by enzyme-linked immunosorbent assay (ELISA) kits. Western blot was used for analysis of the mitogen-activated protein kinase (MAPK) pathways and COX-2 and iNOS protein expression. Topical SP treatment reduced serum IgE and IgG2a levels and suppressed COX-2 and iNOS expression via blocked mitogen-activated protein kinase (MAPK) pathways in DNCB-induced AD-like lesions. Histopathological examination revealed that SP reduced epidermal thickness and collagen accumulation and inhibited mast cells and eosinophils infiltration into the AD-like lesions skin. These results indicate that SP may protect against AD skin lesions through inhibited MAPK signaling pathways and may diminish the infiltration of inflammatory cells to block allergic inflammation.

Download Full-text

Platyphyllenone Induces Autophagy and Apoptosis by Modulating the AKT and JNK Mitogen-Activated Protein Kinase Pathways in Oral Cancer Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22084211 ◽

2021 ◽

Vol 22 (8) ◽

pp. 4211

Author(s):

Yen-Tze Liu ◽

Hsin-Yu Ho ◽

Chia-Chieh Lin ◽

Yi-Ching Chuang ◽

Yu-Sheng Lo ◽

...

Keyword(s):

Protein Kinase ◽

Oral Cancer ◽

Protein Expression ◽

Caspase 3 ◽

Therapeutic Option ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Dependent Manner ◽

Cancer Proliferation ◽

Mitogen Activated Protein

Platyphyllenone is a type of diarylheptanoid that exhibits anti-inflammatory and chemoprotective effects. However, its effect on oral cancer remains unclear. In this study, we investigated whether platyphyllenone can promote apoptosis and autophagy in SCC-9 and SCC-47 cells. We found that it dose-dependently promoted the cleavage of PARP; caspase-3, -8, and -9 protein expression; and also led to cell cycle arrest at the G2/M phase. Platyphyllenone up-regulated LC3-II and p62 protein expression in both SCC-9 and SCC-47 cell lines, implying that it can induce autophagy. Furthermore, the results demonstrated that platyphyllenone significantly decreased p-AKT and increased p-JNK1/2 mitogen-activated protein kinase (MAPK) signaling pathway in a dose-dependent manner. The specific inhibitors of p-JNK1/2 also reduced platyphyllenone-induced cleavage of PARP, caspase-3, and caspase -8, LC3-II and p62 protein expression. These findings are the first to demonstrate that platyphyllenone can induce both autophagy and apoptosis in oral cancers, and it is expected to provide a therapeutic option as a chemopreventive agent against oral cancer proliferation.

Download Full-text