기초 : 톡소포자충 감염 신경교종세포의 Mitogen-Activated Protein Kinase Pathway 활성화

박병현 ( Byung Hyun Park ),이현성 ( Hyun Sung Lee ),이종수 ( Jong Soo Lee ),양승환 ( Seung Hwan Yang ),신대환 ( Dae Whan Shin ),이영하 ( Young Ha Lee ) 대한뇌종양학회 2006 대한뇌종양학회지 Vol.5 No.2

Objective:Gliomas are the most common primary brain tumors in adults. Toxoplasma gondii is an obligate intracellular parasite with a high affinity for brain cells. Mitogen-activated protein kinases(MAPKs) are known to regulate the host cell invasion, although little is known regarding MAPK signaling in Toxoplasma-infected glioma cells. Methods:U87MG and U373MG cells were infected with tachyzoites of the RH strain of T. gondii, and the kinetics of MAPK activation were determined by immunoblotting. We also determined the effects of MAPK inhibitors on glioma cell growth and T. gondii replication using the 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetraterazolium bromide(MTT; Sigma) method and [3H]-uracil incorporation assays. Results:There were no significant differences in the levels of unphosphorylated MAPK at different incubation times in the Toxoplasma-infected U373MG cells. T. gondii induced the activation of extracellular signal-regulated kinase(ERK) 1/2, p30MAPK, and c-Jun N-terminal kinase(JNK)1/2 within 30 minutes of infection, and showed differential kinetics of activation for each MAPK. The activation of ERK1/2, p38MAPK, and JNK1/2 in Toxoplasma-infected glioma cells was blocked by PD98059, SB202190, and SB203580, respectively. T. gondii replication was inhibited in a dose-dependent manner by the addition of MAPK inhibitors to Toxoplasma-infected glioma cells. Conclusion:Infection with T. gondii induces the activation of MAPKs in glioma cells, and this activation can be blocked by the addition of kinase-specific inhibitors. These results suggest that the MAPK pathway plays a role in the invasion of glioma cells with T. gondii.

Roles of Mitogen-Activated Protein Kinases in Osteoclast Biology

Lee, Kyunghee,Seo, Incheol,Choi, Mun Hwan,Jeong, Daewon MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.10

<P>Bone undergoes continuous remodeling, which is homeostatically regulated by concerted communication between bone-forming osteoblasts and bone-degrading osteoclasts. Multinucleated giant osteoclasts are the only specialized cells that degrade or resorb the organic and inorganic bone components. They secrete proteases (e.g., cathepsin K) that degrade the organic collagenous matrix and establish localized acidosis at the bone-resorbing site through proton-pumping to facilitate the dissolution of inorganic mineral. Osteoporosis, the most common bone disease, is caused by excessive bone resorption, highlighting the crucial role of osteoclasts in intact bone remodeling. Signaling mediated by mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38, has been recognized to be critical for normal osteoclast differentiation and activation. Various exogenous (e.g., toll-like receptor agonists) and endogenous (e.g., growth factors and inflammatory cytokines) stimuli contribute to determining whether MAPKs positively or negatively regulate osteoclast adhesion, migration, fusion and survival, and osteoclastic bone resorption. In this review, we delineate the unique roles of MAPKs in osteoclast metabolism and provide an overview of the upstream regulators that activate or inhibit MAPKs and their downstream targets. Furthermore, we discuss the current knowledge about the differential kinetics of ERK, JNK, and p38, and the crosstalk between MAPKs in osteoclast metabolism.</P>

Increased interaction between heat shock protein 27 and mitogen-activated protein kinase (p38 and extracellular signal-regulated kinase) in pre-eclamptic placentas

Shin, Jeong-Kyu,Jeong, Young-Taek,Jo, Hyun-Cheol,Kang, Min-Young,Chang, In-Suk,Baek, Jong-Chul,Park, Ji-Kwon,Lee, Soon-Ae,Lee, Jong-Hak,Choi, Wan-Sung,Paik, Won-Young Blackwell Publishing Asia 2009 JOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH -TO Vol.35 No.5

<P>Abstract</P><P>Aims: </P><P>Heat shock protein 27 (Hsp27) is a well-known stress response protein that is characterized by its phosphorylative capacity. Hsp27 becomes phosphorylated in response to various stimuli through interaction with several different kinases. The purpose of this study was to evaluate the interaction between Hsp27 and mitogen-activated protein kinase (MAPK) (p38, extracellular signal-regulated kinase [ERK], and c-Jun N-terminal kinase) in the human placenta derived from patients with pre-eclampsia.</P><P>Methods: </P><P>Western blot analysis was used to examine the levels of expression of Hsp27 and MAPK (p38, ERK, and c-Jun N-terminal kinase). Immunoprecipitation analysis was used to determine the interaction between Hsp27 and MAPK (p38 and ERK).</P><P>Results: </P><P>Western blotting analysis and immunohistochemistry showed that the expression of Hsp27 and p-Hsp27 in the placental tissues of the pre-eclampsia group were significantly higher than that in the normal pregnancy group. Immunoprecipitation analysis showed that the interaction between Hsp27 and MAPK (p38 and ERK) was significantly increased in the pre-eclamptic placenta tissues.</P><P>Conclusion: </P><P>The interaction between Hsp27 and MAPK was increased, suggesting that phosphorylation of Hsp27 might be induced by p38 and ERK in placentas from patients with pre-eclampsia.</P>

Bone Morphogenetic Protein 2-induced MAPKs Activation Is Independent of the Smad1/5 Activation

Jun, Ji Hae,Ryoo, Hyun-Mo,Woo, Kyung Mi,Kim, Gwan-Shik,Baek, Jeong-Hwa KOREAN ACADAMY OF ORAL BIOLOGY 2009 International Journal of Oral Biology Vol.34 No.2

Bone morphogenetic protein (BMP) 2 is a potent osteogenic factor. Although both Smad1/5 and mitogen-activated protein kinases (MAPKs) are activated by BMP2, the hierarchical relationship between them is unclear. In this study, we examined if BMP2-stimu1ated MAPK activation is regulated by Smad1/5 or vice versa. When C2C12 cells were treated with BMP2, the activation of extracellular signal-regulated kinasu (ERK), p38 MAPK and c-Jun-N-terminal kinase was evident within 5 min. The knockdown of both Smad1 and Smad5 by small interfering RNA did not affect the activation of these MAPKs. In addition, neither the overexpression of Smad1 nor Smad5 induced ERK activation. When ERK activation was induced by constitutively active MEK1 expression, the protein level and activation of Smad1 increased. Furthermore, the inhibition of constitutively active BMP receptor type IB-induced ERK activation significantly suppressed Smad1 activation. These results indicate that Smad1/5 activation is not necessary for BMP2-induced MAPK activation and also that ERK positively regulates Smad1 activation.

Carvedilol이 배양된 사람 혈관 평활근 세포의 증식과 그에 관여하는 세포내 신호전달계에 미치는 영향

박제현,하헌주,오재원,김명수,서지연,김혜진,박기일,김유선 대한혈관외과학회 2002 Vascular Specialist International Vol.18 No.1

장기이식 후 만성거부반응이나 혈관손상 후 재협착 등의 복원과정(remodeling) 그리고 동맥경화증의 병태생리는 비슷하여 물리적 손상이나 면역학적 또는 비면역학적 원인에 의해서 혈관내피의 손상이 발생하면 혈관 평활근 세포의 증식과 이동이 항진되며 세포 외 기질이 과다 생산되어 혈관의 내막증식과 섬유화가 초래된다. 이러한 병태 생리과정을 효과적으로 제어하는 방법은 매우 제한적으로 여러 약제를 사용하여 다양한 cytokine과 성장인자의 생성과 빈혈을 억제함으로써 혈관 병변과 재협착을 억제하고자 하는 시도가 있어왔으나 임상에서 사용할 정도로 그 효과가 확연하게 밝혀진 제제는 아직까지 없다)1-3). 연구자들은 최근에 항고혈압제로 사용중인 carvedilol 제제가 백서의 혈관 평활근세포의 증식과 이동을 효과적으로 억제함을 관찰하여 보고한 바 있다(4-5). 아드레날린성 β억제제로 개발된 carvedilol은 아드레날린성 α억제제 및 항산화제 등 다양한 기능을 가진 제제로서(6) 다양한 혈관 병변을 가진 환자나 신장이식환자에서 항고혈압제제의 복용이 필요한 상황을 고려해 보면 carvedilol의 투여는 혈압 강하효과 이외에도 혈관병변의 예방과 치료에 효과가 있을 것으로 사료된다. 세포의 성장과 증식은 이를 촉진하거나 억제하는 유전자 발현과 단백질 합성에 의해 이루어지며 정상 생리 상태에서는 유기적으로 잘 조절되는 신호변환기전에 의해서 조절된다. 성장인자는 세포막에 있는 수용체와 결합하여 그 수용체를 활성화시킨 후 신호변환기전을 경유하여 핵내의 유전자 발현을 조절한다. 특히, mitogen-activated protein kinases (MAPK)는 세포질에 존재하는 단백질의 인산화 효소로서 extra-cellular-regulatory protein kinase (ERK), c-jun N-terminal kinase (JNK), p38 MAPK의 세 가지 형태가 있으며, 각각 세포외부의 각종 자극 인자들에 의해 연속적으로 활성화되어 전사조절인자를 활성화한다(7-11). 최근의 보고에 따르면 MAPK나 전사조절인자의 활성조절에는 활성산소족(reactive oxygen species)과 이들에 의해 유도되는 세포내 산화-환원 상태의 변화가 상당부분 관여하는 것으로 알려져 있다(7,12,13). 따라서, 본 연구자들은 carvedilol 제제가 인체 유래 혈관 평활근세포의 증식에 미치는 영향과 이에 관여하는 신호 전달계 중 활성산소족생성과 MAPK의 활성화에 미치는 영향을 규명하기 위하여 본 실험을 실시하였다. Purpose: Vascular smooth muscle cells (VSMSs) migration and proliferation play important roles in transplant vascular sclerosis and restenosis afer balloon vascular injury. The antiproliterative and anti-migratory effects of carvedilol (CA), a unique α-and β-blocking anti-hypertensive drug, on the VSMCs were confirmed previously. Since reactive oxygen species (ROS) and mitogen-activated protein kinases (MAPK) family play important roles in proliferation of VSMCs, the present study examined the effects of CA on intracellular ROS generation, activation of ERK 1/2 and p38 MAPK, and proliferation of VSMCs were cultured with RPMI-1640 containing 10% fetal bovine serum. Near confluent VSMCs were incubated with serum-free media for 48 hours to arrest and synchronize the cell growth. CA was administered 1 hour before the addition of PDGF. 5-(and-6)-chloromethy-2',7'-dichlorodihydrofluo-rescein (DCF)-sensitive intracellular ROS was detected by FACS. Activations of ERK 1/2 and p38 MAPK were measured by Western blot analysis, Proliferation of VSMCs was assessed by [^3H]-thymidine incorporation. Results: PDGF at 10 ??/㎖, which induced human VSMCs proliferation, rapidly increased intracellular ROS by 1.6-fold (P < 0.01), ERK 1/2 activation by 2.1-fold (P < 0.01), and p38 MAPK activation by 1.9-fold (P < 0.01), respectively, as compared to the control. CA 1 and 10μM effectively inhibited PDGF-induced human VSMCs proliferation. CA also effectively inhibited PDGF-induced intracellular

The Mitogen-Activated Protein Kinase Signal Transduction Pathways in Alternaria Species

Houjuan Xu,Xiaoxue Xu,Yujun Wang,비벡바지파이,Lisha Huang,Yongfang Chen,백광현 한국식물병리학회 2012 Plant Pathology Journal Vol.28 No.3

Mitogen-activated protein kinase (MAPK) cascades are conserved signaling modules in the eukaryotic cells. They are involved in many major cell processes in fungi such as stress responses, vegetative growth, pathogenicity,secondary metabolism and cell wall integrity. In this review, we summarized the advances of research on the MAPK signaling pathways in Alternaria species. As major phytopathogenic fungi, Alternaria species reduce crop production. In contrast to the five MAPK pathways known in yeast, only three MAPK pathways as Fus3/Kss1-type, Hog1-type, and Slt2-type have been characterized in Alternaria. The Fus3/Kss1-type MAPK pathway participates in regulation of vegetative growth,conidiation, production of some cell-wall-degrading enzymes and pathogenicity. The Hog1-type pathway is involved in osmotic and oxidative stress, fungicides susceptibility and pathogenicity. The Slt2-type MAP kinases play an important role on maintaining cell wall integrity, pathogenicity and conidiation. Although recent advances on the MAPK pathways in Alternaria spp. reveal many important features on the pathogenicity,there are many unsolved problems regarding to the unknown MAP kinase cascade components and network among other major signal transduction. Considering the economic loss induced by Alternaria spp., more researches on the MAPK pathways will need to control the Alternaria diseases.

Rhythmic Expression of Mitogen Activated Protein Kinase Activity in Rice

Kudupudi Prabhakara Rao,Gubbala Vani,Kundan Kumar,Alok Krishna Sinha 한국분자세포생물학회 2009 Molecules and cells Vol.28 No.5

Mitogen activated protein kinase (MAPK) are known to get activated during various stress signals and transduce the message from the cell membrane to the nucleus for ap-propriate cellular reorganization. Though, a certain basal activity of MAPK is often observed in the control plants. Prolonged exposure of rice plants to lowered or elevated temperature exhibited a rhythm in the activation of MAPKs. We analyzed existence of a possible endogenous rhythm in the activity of MAPKs in rice plants. The plants growing at constant temperature entrained in 16/8 h day-night cycle showed diurnal rhythm in activity. When the activation of MAPK was tested under continuous conditions by shifting plants to continuous darkness for a period of 72 h, the periodic rhythm persisted and followed a circadian pattern. Analysis of the transcripts of group A, B and C members of MAPKs under above conditions by quantitative real time PCR revealed that the members of group C exhibit periodic rhythm. Our data indicates that the MAP kinase activity in rice follows rhythmic expression in a circadian manner.

The Mitogen-Activated Protein Kinase Signal Transduction Pathways in Alternaria Species

Xu, Houjuan,Xu, Xiaoxue,Wang, Yu-Jun,Bajpai, Vivek K.,Huang, Lisha,Chen, Yongfang,Baek, Kwang-Hyun The Korean Society of Plant Pathology 2012 Plant Pathology Journal Vol.28 No.3

Mitogen-activated protein kinase (MAPK) cascades are conserved signaling modules in the eukaryotic cells. They are involved in many major cell processes in fungi such as stress responses, vegetative growth, pathogenicity, secondary metabolism and cell wall integrity. In this review, we summarized the advances of research on the MAPK signaling pathways in Alternaria species. As major phytopathogenic fungi, Alternaria species reduce crop production. In contrast to the five MAPK pathways known in yeast, only three MAPK pathways as Fus3/Kss1-type, Hog1-type, and Slt2-type have been characterized in Alternaria. The Fus3/Kss1-type MAPK pathway participates in regulation of vegetative growth, conidiation, production of some cell-wall-degrading enzymes and pathogenicity. The Hog1-type pathway is involved in osmotic and oxidative stress, fungicides susceptibility and pathogenicity. The Slt2-type MAP kinases play an important role on maintaining cell wall integrity, pathogenicity and conidiation. Although recent advances on the MAPK pathways in Alternaria spp. reveal many important features on the pathogenicity, there are many unsolved problems regarding to the unknown MAP kinase cascade components and network among other major signal transduction. Considering the economic loss induced by Alternaria spp., more researches on the MAPK pathways will need to control the Alternaria diseases.

Gene structure, molecular characterization and transcriptional expression of two p38 isoforms (MAPK11 and MAPK14) from rock bream (Oplegnathus fasciatus)

Umasuthan, N.,Bathige, S.D.N.K.,Noh, J.K.,Lee, J. Academic Press 2015 FISH AND SHELLFISH IMMUNOLOGY Vol.47 No.1

The p38 kinases are one of the four subgroups of mitogen-activated protein kinase (MAPK) superfamily which are involved in the innate immunity. The p38 subfamily that includes four members namely p38α (MAPK14), p38β (MAPK11), p38γ (MAPK12) and p38δ (MAPK13), regulates the activation of several transcription factors. In this study, a p38β (OfMAPK11) homolog and a p38α (OfMAPK14) homolog of Oplegnathus fasciatus were identified at genomic level. Results clearly showed that both MAPK11 and MAPK14 are well-conserved at both genomic structural- and amino acid (aa)-levels. Genomic sequences of OfMAPK11 (~15.6 kb) and OfMAPK14 (~13.4 kb) had 12 exons. A comparison of exon-intron structural arrangement of these genes from different vertebrate lineages indicated that all the exon lengths are highly conserved, except their terminal exons. Full-length cDNAs of OfMAPK11 (3957 bp) and OfMAPK14 (2504 bp) encoded corresponding proteins of 361 aa and 360 aa, respectively. Both OfMAPK proteins harbored a Ser/Thr protein kinases catalytic domain (S_TKc domain) which includes an activation loop with a dual phosphorylation site (TGY motif) and several specific-binding sites for ATP and substrates. Molecular modeling of the activation loop and substrate binding sites of rock bream MAPKs revealed the conservation of crucial residues and their orientation in 3D space. Transcripts of OfMAPKs were ubiquitously detected in eleven tissues examined, however at different levels. The modulation of OfMAPKs' transcription upon pathogen-associated molecular patterns (PAMPs: flagellin, lipopolysaccharide and poly I:C) and pathogens (Edwardsiella tarda, Streptococcus iniae and rock bream iridovirus) was investigated. Among the seven examined tissues, the flagellin-challenge upregulated the mRNA level of both OfMAPKs in the head kidney. Meanwhile, modulation of OfMAPK mRNA expression in the liver upon other immune-challenges varied in a time-dependent manner. Collectively, these results suggest that OfMAPKs are true members of p38 subfamily, which might be induced by different immune stimuli.

Selective modulation of the glucocorticoid receptor can distinguish between transrepression of NF-κB and AP-1

De Bosscher, Karolien,Beck, Ilse M.,Dejager, Lien,Bougarne, Nadia,Gaigneaux, Anthoula,Chateauvieux, Sé,bastien,Ratman, Dariusz,Bracke, Marc,Tavernier, Jan,Vanden Berghe, Wim,Libert, Claude,Diede Springer Basel 2014 Cellular and molecular life sciences Vol.71 No.1

<P>Glucocorticoids (GCs) block inflammation via interference of the liganded glucocorticoid receptor (GR) with the activity of pro-inflammatory transcription factors NF-κB and AP-1, a mechanism known as transrepression. This mechanism is believed to involve the activity of GR monomers. Here, we explored how the GR monomer-favoring Compound A (CpdA) affects AP-1 activation and activity. Our results demonstrate that non-steroidal CpdA, unlike classic steroidal GCs, blocks NF-κB- but not AP-1-driven gene expression. CpdA rather sustains AP-1-driven gene expression, a result which could mechanistically be explained by the failure of CpdA to block upstream JNK kinase activation and concomitantly also phosphorylation of c-Jun. In concordance and in contrast to DEX, CpdA maintained the expression of the activated AP-1 target gene <I>c</I>-<I>jun</I>, as well as the production of the c-Jun protein. As for the underlying mechanism, GR is a necessary intermediate in the CpdA-mediated gene expression of AP-1-regulated genes, but seems to be superfluous to CpdA-mediated JNK phosphorylation prolongation. The latter phenomenon concurs with the inability of CpdA to stimulate DUSP1 gene expression. ChIP analysis demonstrates that DEX-activated GR, but not CpdA-activated GR, is recruited to AP-1-driven promoters. Furthermore, in mice we observed that CpdA instigates a strong enhancement of TNF-induced AP-1-driven gene expression. Finally, we demonstrate that this phenomenon coincides with an increased sensitivity towards TNF lethality, and implicate again a role for JNK2. In conclusion, our data support the hypothesis that a ligand-induced differential conformation of GR yields a different transcription factor cross-talk profile.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (doi:10.1007/s00018-013-1367-4) contains supplementary material, which is available to authorized users.</P>