흰쥐 뇌에서 전기경련 충격에 의한 CREB 인산화의 발달단계에 따른 변화

강웅구,정희연,안용민,정선주,전송희,박주배,조수철,김용식 大韓神經精神醫學會 1999 신경정신의학 Vol.38 No.3

연구목적 : 이 논문에서는 뇌에서 전기경련 충격(Electroconvulsive shock. ECS)에 의한 전사인자 CREB의 인산화를 발달단계에 따라 알아봄으로써 정신장애의 신경발달학적 이해를 위한 생물학적 기반지식을 얻고자 하였다. 방 법 : 생후 7. 14. 21일 및 성년 흰쥐에 ECS를 가하고 시간별로 해마 및 소뇌 조직을 얻어 CREB의 인산화를 알아보기 위해 특이 항체로 면역블롯을 실시하였다. 결 과 : 발달단계에 따라 해마에서는 CREB이 감소한 반면 소뇌에서는 CREB이 증가하였다. 기저상태의 CREB 인산화는 해마와 소뇌에서 생후 7일에 비해 14일 이후 증가하였는데, 소뇌의 경우 CREB의 양증가와 비례하였다. ECS 후 CREB인산화 증가는 해마에서는 생후 21일 이후에 나타났으나, 소뇌에서는 생후 7일은 물론 성년에서도 나타나지 않았다. 결 론 : CREB 매개 신호전달은 발달단계별 및 조직별 활성의 차이를 나타내었으며 해마에서는 생후 21일 이후 ECS에 의해 활성화되었지만, 소뇌에서는 그렇지 않았다. 해마에서 ECS에 의한 CREB 인산화 증가는 c-fos 유전자의 발현과 관계있으리라고 생각되지만, CREB의 Ser-133 인산화 만으로는 발달단계 및 조직에 따른 c-fos 발현의 특이성을 설명할 수 없었다. Objectives : In order to understand the biological basis of neurodevelopmental perspectives of mental disorders, the authors investigated the developmental and regional changes in the phosphorylation of the transcription factor CREB following the electroconvulsive shock(ECS) in rat brain. Methods : Rats of various age groups(7, 14, 21 days postnatal and adults) were given ECS and their hippocampi and cerebella were dissected at specified time points. The content of CREB and phosphorylated CREB were measured by immunoblot analysis. Results : The amount of CREB increased in the hippocampus and decreased in the cerebellum according to the age. Baseline levels of CREB phosphorylation in both tissues were increased from postnatal 14 days, and it was proportional to the amount of CREB protein in the cerebellum. In the hippocampus, ECS increased the phosphorylation of CREB at postnatal 21 days, but in the cerebellum, ECS did not increased the phosphorylation of CREB in any age group. Conclusion : CREB mediated signal transduction pathways showed developmental and tissue-specific changes. ECS increased the phosphorylation of CREB in the hippocampus by postnatal 21 days, but not in the cerebellum. CREB activation is supposed to be related with the induction of c-fos after ECS in the hippocampus. However, the Ser-133 phosphorylation of CREB could not completely explain the developmental and tissue specificity of c-fos induction.

A Functional Role for CREB as a Positive Regulator of Memory Formation and LTP

Satoshi Kida 한국뇌신경과학회 2012 Experimental Neurobiology Vol.21 No.4

cAMP response element-binding protein (CREB), a transcription factor, has been shown to play a central role in memory formation, and its involvement in this process has been investigated using a wide range of animal models, from nematodes to higher animals. Various CREB mutant mice have been developed and investigated. Several types of mutant mice with loss of CREB function have impaired memory formation and long-term potentiation (LTP), suggesting that CREB plays essential roles in these processes. To characterize the roles of CREB in memory formation and LTP further, mutant mice displaying gain of CREB function have been generated and analyzed. Importantly, CREB-DIEDML mice and CREB-Y134F mice showed enhanced memory formation, whereas CREB-VP16 mice displayed a lowered threshold of long-lasting LTP (L-LTP) induction, strongly suggesting that CREB functions as a positive regulator of memory formation and LTP. In this review, I focus on the effects of the genetic activation of CREB in LTP and memory formation and summarize previous findings.

The SMILE transcriptional corepressor inhibits cAMP response element–binding protein (CREB)–mediated transactivation of gluconeogenic genes

Lee, Ji-Min,Han, Hye-Sook,Jung, Yoon Seok,Harris, Robert A.,Koo, Seung-Hoi,Choi, Hueng-Sik American Society for Biochemistry and Molecular Bi 2018 The Journal of biological chemistry Vol.293 No.34

<P>Under fasting conditions, activation of several hepatic genes sets the stage for gluconeogenesis in the liver. cAMP response element-binding protein (CREB), CREB-regulated transcription coactivator 2 (CRTC2), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1 alpha) are essential for this transcriptional induction of gluconeogenic genes. PGC-1insight that may help inform potential therapeutic approaches targeting PGC-l alpha-mediated regulation of hepatic glucose metabolism. induction is mediated by activation of a CREB/CRTC2 signaling complex, and recent findings have revealed that small heterodimer partner-interacting leucine zipper protein (SMILE), a member of the CREB/ATF family of basic region-leucine zipper (bZIP) transcription factors, is an insulin-inducible corepressor that decreases PGC-1 alpha expression and abrogates its stimulatory effect on hepatic gluconeogenesis. However, the molecular mechanism whereby SMILE suppresses PGC-1a expression is unknown. Here, we investigated SMII.E's effects on the CREB/CRTC2 signaling pathway and glucose metabolism. We found that SMILE significantly inhibits CREB/ CRTC2-induced PGC-1 alpha expression by interacting with and disrupting the CREB/CRTC2 complex. Consequently, SMILE decreased PGC-1 alpha-induced hepatic gluconeogenic gene expression. Furthermore, SMILE inhibited CREB/CRTC2-induced phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) gene expression by directly repressing the expression of these genes and by indirectly inhibiting the expression of PGC-1 alpha via CREB/CRTC2 repression. Indeed, enhanced gluconeogenesis and circulating blood glucose levels in mice injected with an adenovirus construct containing a constitutively active CRTC2 variant (CRTC2-S171A) were significantly reduced by WT SMILE, but not by leucine zipper-mutated SMILE. These results reveal that SMILE represses CREB/CRTC2-induced PGC-1 alpha expression, an insight that may help inform potential therapeutic approaches targeting PGC-1 alpha-mediated regulation of hepatic glucose metabolism.</P>

Clenbuterol Inhibits SREBP-1c Expression by Activating CREB1

Zhou, Lei,Li, Yixing,Nie, Tao,Feng, Shengqiu,Yuan, Jihong,Chen, Huaping,Yang, Zaiqing Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.4

As a $\beta_2$-adrenergic agonist, clenbuterol decreases body fat, but the molecular mechanism underlying this process is unclear. In the present study, we treated 293T and L-02 cells with clenbuterol and found that clenbuterol downregulates SREBP-1c expression and upregulates CREB1 expression. Considering SREBP-1c has the function of regulating the transcription of several lipogenic enzymes, we considered that the downregulation of SREBP-1c is responsible for body fat reduction by clenbuterol. Many previous studies have found that clenbuterol markedly increases intracellular cAMP levels, therefore, we also investigated whether CREB1 is involved in this process. The data from our experiments indicate that CREB1 overexpression inhibits SREBP-1c transcription, and that this action is antagonized by CREB2, a competitive inhibitor of CREB1. Furthermore, since PPARs are able to repress SREBP-1c transcription, we investigated whether clenbuterol and CREB1 function via a pathway involving PPAR activation. However, our results showed that clenbuterol or CREB1 overexpression suppressed PPARs transcription in 293T and L-02 cells, which suggested that they impair SREBP-1c expression in other ways.

Antidepressant effects of aqueous extract of saffron and its effects on CREB, P-CREB, BDNF, and VGF proteins in rat cerebellum

Najmeh Asrari,Rezvan Yazdian-Robati,Khalil Abnous,Bibi Marjan Razavi,Mrazieh Rashednia,Faezeh Vahdati Hasani,Hossein Hosseinzadeh 대한약침학회 2018 Journal of pharmacopuncture Vol.21 No.1

Objective: The role of BDNF (brain-derived neurotrophic factor), CREB (cAMP response element binding) and VGF neuropeptide has been proved in antidepressant activity of long term saffron administration in the rat hippocampus. In this study we evaluated the role of these proteins in antidepressant activity of saffron in long term administration in the rat cerebellum. Methods: Saffron aqueous extract (40 and 80 mg/kg/ day) and imipramine (10 mg/kg/day) were administered intraperitoneally for 21 days to rats. At the end of experiment, animals were sacrificed and cerebellums were separated. The protein levels of BDNF, VGF, CREB and P- CREB in the rat cerebellum were evaluated using western blot analysis. Results: Saffron aqueous extract (80mg/kg/day) caused significant increase in protein level of P-CREB in long term treatment in the rat cerebellum. The increases in the protein levels of VGF, CREB and BDNF were not significant. Conclusion: In summary, our results showed that antidepressant effect of saffron in rat cerebellum might be due to the enhanced phosphorylation of CREB.

Antidepressant effects of aqueous extract of saffron and its effects on CREB, P-CREB, BDNF, and VGF proteins in rat cerebellum

Asrari, Najmeh,Yazdian-Robati, Rezvan,Abnous, Khalil,Razavi, BiBi Marjan,Rashednia, Mrazieh,Hasani, Faezeh Vahdati,Hosseinzadeh, Hossein KOREAN PHARMACOPUNCTURE INSTITUTE 2018 Journal of pharmacopuncture Vol.21 No.1

Objective: The role of BDNF (brain-derived neurotrophic factor), CREB (cAMP response element binding) and VGF neuropeptide has been proved in antidepressant activity of long term saffron administration in the rat hippocampus. In this study we evaluated the role of these proteins in antidepressant activity of saffron in long term administration in the rat cerebellum. Methods: Saffron aqueous extract (40 and 80 mg/kg/day) and imipramine (10 mg/kg/day) were administered intraperitoneally for 21 days to rats. At the end of experiment, animals were sacrificed and cerebellums were separated. The protein levels of BDNF, VGF, CREB and P- CREB in the rat cerebellum were evaluated using western blot analysis. Results: Saffron aqueous extract (80mg/kg/day) caused significant increase in protein level of P-CREB in long term treatment in the rat cerebellum. The increases in the protein levels of VGF, CREB and BDNF were not significant. Conclusion: In summary, our results showed that antidepressant effect of saffron in rat cerebellum might be due to the enhanced phosphorylation of CREB.

P-92 The Role of PAK1/CREB axis in Carcinogenesis of Squamous Cell Lung Cancer

정재헌,김윤성,윤성훈 대한결핵 및 호흡기학회 2017 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.124 No.-

Background: The p21-activated Ser/Thr kinase 1 (PAK1) kinase has an essential role in tumorigenesis in many cancers, but whether PAK1 expression correlates with NSCLC tumorigenesis and its mechanism is still poorly understood. We hypothesized that PAK1 could modulate the cyclic AMP response element binding (CREB) protein expression and its activity, which have a central role in lung carcinogenesis as a regulatory factor in NSCLC. Methods: Clinical data were retrospectively reviewed in 60 patients with squamous cell lung cancer from the tissue microarray. Western blotting, immunohistochemical analysis, and TUNEL were performed to study for the expression and interaction of PAK1 and CREB. Results: We showed that in squamous lung cancer tissues, PAK1 was upregulated (3-fold to 4-fold, P=0.015), tyrosine phosphorylated (3-fold, P<0.01), and translocated to the nucleus. PAK1 siRNA-transfected downregulation induced apoptosis in squamous carcinoma cells by cleavage and activation of downstream effector caspases. Consistent with the results from Western blotting, apoptotic cell death was detected by TUNEL assay. Overexpression of PAK1 was positively correlated with CREB activity (r=0.69, P<0.05). Notably, PAK1 blockade did not change the level of CREB but modify CREB activity (P<0.005). We performed Western blotting from over-expression plasmid transfected cells and found that PAK1 phosphorylates CREB at Ser133. Conclusion: Our findings offered evidence that PAK1 phosphorylates CREB in lung squamous carcinomas, with implications for regulation of PAK1 as a potential therapeutic target of squamous lung cancer treatment.

운동에 의한 골격근 내 PGC-1α전사 조절 기전

김상현,고진호,김필상,김기진 한국운동생리학회(구 한국운동과학회) 2013 운동과학 Vol.22 No.3

본 연구는 PGC-1α의 promoter에 존재하는 CRE 서열에 결합하여 PGC-1α의 전사를 유도하는 ATF2와 CREB의 활성화를 중심으로 운동 시 골격근 내 PGC-1α의 전사가 어떻게 이루어지는지 알아보았다. 이를 위해 luciferase reporter system을 이용하여 골격근 내 ATF2와 CREB의 인산화 그리고 PGC-1α의 전사를 확인하기 위해 생쥐의 골격근세포에 clenbuterol(β2-adrenergic agonist)과 anisomycin(p38 MAPK activator)을 처치하였다. 또한 흰쥐 12마리를 무작위로 두 그룹(Sedentary와 Exercise)으로 분류한 후 운동 그룹은 30분간 수영(2% weight/body weight, 30 min)을 실시하였으며, 운동직후 CREB과 ATF2의 인산화 정도를 확인하기 위해 골격근을 적출하였다. 그 결과 clenbuterol 처치는 CREB의 인산화를 증가시켰으며, anisomycin의 처치는 ATF2의 인산화를 증가시켰다. 그러나 운동에 의해서는 ATF2만 인산화가 증가하였다. 그리고 PGC-1α의 전사 또한 ATF2의 인산화에 의해서만 유도되었으며, DN-ATF2에 의해서는 PGC-1α의 전사가 완벽히 억제되었다. 이러한 결과는 운동 시 골격근 내 p38 MAPK 기전의 활성에 의한 ATF2 인산화가 PGC-1α의 전사를 유도함을 나타낸다. This study determines which transcriptional factors, activating transcription factor 2 (ATF2) and/or cAMP responsive element binding protein (CREB), bind to and activate cAMP response element (CRE) transcriptional binding site of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) promoter during exercise. C2C12 cells were treated clenbuterol(β2-adrenergic agonist) and anisomycin (p38 MAPK activator) to determine weather p-CREB and/or p-ATF2 can activate PGC-1α promoter transcriptional activities using luciferase reporter system. Twelve rats were randomly assigned into 2 groups (Sedentary and Exercise). Exercise group was trained by using a 30min swimming with tail load equivalent to 2 % of animal`s own body weight. Muscle samples were harvested immediately after exercise. Clenbuterol and anisomycin increased phosphorylation of CREB and ATF2 respectively in C2C12. The ATF2 phosphorylation was increased in the exercise group compared with sedentary group, but CREB phosphorylation was not changed after exercise. PGC-1α transcription activities were enhanced by ATF2 activation, but not in CREB activated condition. Induction of DN-ATF2 completely blocked PGC-1α promoter activity by p-ATF2. These findings suggest that ATF2 phosphorylation by p38 MAPK system induce PGC-1α transcription in exercised muscle.

구연 : 간암세포주 HepG2와 SNU182에서 AP 1과 CREB의 활성에 미치는 PD98059의 영향

김성우,조승철,한성희,감창우,박재일,손주현,이동후,기춘석 대한간학회 2003 Clinical and Molecular Hepatology(대한간학회지) Vol.9 No.3(S)

배경/목적: Transcriptional transactivator인 activator protein 1 (AP 1)과 cyclic AMP response element binding (CREB)은 신호전달체계의 자극을 받아 세포 생존과 증식에 필수적인 것으로 알려져 있다. PD98059는 mitogen-activated protein kinase (MAPK)의 activator인 MAP kinase / extracellular signal-regulated kinase (ERK) kinase (MEK)을 억제한다. 본 연구에서는 PD98059가 간암세포주 HepG2와 SNU182에서 AP 1과 CREB의 발현에 미치는 영향을 추구하였다. 대상과 방법: 간암세포주 HepG2와 SNU182의 계대 배양에서 PD98059를 0, 5, 10, 50, 75 mol씩 첨가한 후 6, 18, 24, 48, 72시간 배양하여 ELISA를 이용한 MTT assay로 성장 정도를 평가하였다. 그리고 세포핵 RNA를 분리한 것을 electrophoretic mobility shift assay (EMSA)로 검색한 것을 5`-[γ-32P] ATP labeled oligonucleotide로 표지시킨 AP 1과 CREB 탐지자로 각각의 발현 정도를 평가하였다. 결과: EMSA 결과 HepG2에서의 AP 1의 활성은 PD98059의 용량(5, 10, 50, 75 mol) 증가와 더불어 감소되었으나 6시간 배양에서는 미약하였고 18시간 24시간 경과 시 현저하였으나 이후 48시간 72시간에서는 용량에 관계없이 그 발현이 회복되었다. SNU182에서 PD98059의 첨가 후 배양 6시간 이후 시간의 경과와 더불어 용량 의존성 AP 1의 발현 감소를 보였다. HepG2에서의 CREB의 발현은 PD98059에 의한 변화를 보이지 않았다. SNU182에서 PD98059의 첨가 후 6시간 8시간 24시간 경과 시 CREB 활성 억제가 미약하였으나 이후 48시간 72시간에서는 용량-의존적으로 CREB 발현 감소를 보였다. MTT assay 결과 HepG2와 SNU182의 성장은 PD98059에 의해 모두 용량 - 의존적으로 시간 경과에 따라 80% 이상까지 억제되었다. 결론: 본 연구의 성적들은 PD98059에 의한 세포신호전달체계의 제어가 B형 간염 바이러스 감염과 무관한 간암세포주인 HepG2에서는 CREB 활성 억제는 못하고 AP 1의 활성 억제와 관련됨을 뒷받침해 준다. 이와는 대조적으로 B형 간염 바이러스 감염과 관련된 SNU182에서의 PD98059에 의한 세포신호전달체계의 제어는 AP 1과 CREB 양자 모두의 발현 억제에 따라 그 성장 감퇴를 초래하는 것으로 판단된다.

CREB/GSK-3β signaling pathway regulates the expression of TR4 orphan nuclear receptor gene

Park, S.S.,Choi, H.,Kim, S.J.,Chang, C.,Kim, E. North-Holland 2016 Molecular and cellular endocrinology Vol.423 No.-

<P>In this study, we show that reduction of glucose concentration increases TR4 expression in 3T3-L1 cells via stimulation of the GSK-3 beta CREB pathway. While GSK-3 beta and CREB increased TR4 expression in 3T3-L1 cells, inhibition of CREB expression or activity resulted in loss of GSK-3 beta-mediated enhancement of TR4 expression. In addition, CREB enhanced murine TR4 promoter activity via direct binding to a cAMP response element located in the promoter, and this CREB effect was further strengthened by GSK-3 beta. Moreover, silencing of TR4 expression by a gene-specific microRNA inhibited CREB-induced lipid accumulation in 3T3-L1 adipocytes, suggesting that TR4 could be a key mediator of CREB-induced lipogenesis. (C) 2016 Elsevier Ireland Ltd. All rights reserved.</P>