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Xiufen Zhang, Yanling Fan, Beibei Liu, Xiaowei Qi, Zijian Guo, Lihua Li
BACKGROUND: Mediator complex 19 (Med19) is a pivotal subunit of the Mediator complex, and its aberrant expression is involved in tumourigenesis. We aimed to explore the mechanism by which Med19 promotes the proliferation of breast cancer. METHODS: Lentivirus-mediated inhibition of Med19, ectopic expression of Med19 and ectopic expression of core-binding factor subunit alpha 2 to translocation 3 (CBFA2T3) were applied in human breast cancer cell lines. Human breast cancer cell proliferation was determined using CCK8 and colony formation assays after lentivirus infection...
August 29, 2016: Breast Cancer: the Journal of the Japanese Breast Cancer Society
Filip Crona, Per-Henrik Holmqvist, Min Tang, Bhumica Singla, Helin Vakifahmetoglu-Norberg, Katrin Fantur, Mattias Mannervik
The Brakeless protein performs many important functions during Drosophila development, but how it controls gene expression is poorly understood. We previously showed that Brakeless can function as a transcriptional co-repressor. In this work, we perform transcriptional profiling of brakeless mutant embryos. Unexpectedly, the majority of affected genes are down-regulated in brakeless mutants. We demonstrate that genomic regions in close proximity to some of these genes are occupied by Brakeless, that over-expression of Brakeless causes a reciprocal effect on expression of these genes, and that Brakeless remains an activator of the genes upon fusion to an activation domain...
November 1, 2015: Developmental Biology
Muriel Boube, Bruno Hudry, Clément Immarigeon, Yannick Carrier, Sandra Bernat-Fabre, Samir Merabet, Yacine Graba, Henri-Marc Bourbon, David L Cribbs
Hox genes in species across the metazoa encode transcription factors (TFs) containing highly-conserved homeodomains that bind target DNA sequences to regulate batteries of developmental target genes. DNA-bound Hox proteins, together with other TF partners, induce an appropriate transcriptional response by RNA Polymerase II (PolII) and its associated general transcription factors. How the evolutionarily conserved Hox TFs interface with this general machinery to generate finely regulated transcriptional responses remains obscure...
May 2014: PLoS Genetics
Concetta Schiano, Amelia Casamassimi, Maria Teresa Vietri, Monica Rienzo, Claudio Napoli
Despite recent treatment advances, an increase in cardiovascular diseases (CVD) mortality is expected for the next years. Mediator (MED) complex plays key roles in eukaryotic gene transcription. Currently, while numerous studies have correlated MED alterations with several diseases, like cancer or neurological disorders, fewer studies have investigated MED role in CVD initiation and progression. The first finding of MED involvement in these pathologies was the correlation of missense mutations in MED13L gene with transposition of the great arteries...
June 2014: Biochimica et Biophysica Acta
Wenxi Yu, Zhichang Zhang, Daliu Min, Qingcheng Yang, Xuefei Du, Lina Tang, Feng Lin, Yuanjue Sun, Hui Zhao, Shuier Zheng, Aina He, Hongtao Li, Yang Yao, Zan Shen
Osteosarcoma (OS) is the most common primary malignant tumour of bone. Nearly 30-40% of OS patients have a poor prognosis despite multimodal treatments. Because the carcinogenesis of OS remains unclear, the identification of new oncogenes that control the tumourigenesis and progression of OS is crucial for developing new therapies. Here, we found that the expression of Mediator of RNA polymerase II transcription subunit 19 (Med19) was increased in OS samples from patients compared to normal bone tissues. Cyclin D1 and cyclin B1 are upregulated in Med19 positive OS tissues...
April 2014: European Journal of Cancer
Marie-Cécile Caillaud, Shuta Asai, Ghanasyam Rallapalli, Sophie Piquerez, Georgina Fabro, Jonathan D G Jones
Plants are continually exposed to pathogen attack but usually remain healthy because they can activate defences upon perception of microbes. However, pathogens have evolved to overcome plant immunity by delivering effectors into the plant cell to attenuate defence, resulting in disease. Recent studies suggest that some effectors may manipulate host transcription, but the specific mechanisms by which such effectors promote susceptibility remain unclear. We study the oomycete downy mildew pathogen of Arabidopsis, Hyaloperonospora arabidopsidis (Hpa), and show here that the nuclear-localized effector HaRxL44 interacts with Mediator subunit 19a (MED19a), resulting in the degradation of MED19a in a proteasome-dependent manner...
December 2013: PLoS Biology
Sarah K Lee, Xu Chen, Liangqun Huang, Laurie A Stargell
The successful synthesis of a transcript by RNA polymerase II (RNAPII) is a multistage process with distinct rate-limiting steps that can vary depending on the particular gene. A growing number of genes in a variety of organisms are regulated at steps after the recruitment of RNAPII. The best-characterized Saccharomyces cerevisiae gene regulated in this manner is CYC1. This gene has high occupancy of RNAPII under non-inducing conditions, defining it as a poised gene. Here, we find that subunits of the head module of Mediator, Med18 and Med20, and Med19 are required for activation of transcription at the CYC1 promoter in response to environmental cues...
December 2013: Nucleic Acids Research
Li-Jun Zhu, Wang-Xiang Yan, Zhong-Wei Chen, Yu Chen, Dan Chen, Tong-Han Zhang, Gui-Qing Liao
BACKGROUND: Mediator complex subunit 19 (Med19) is a critical subunit of the mediator complex that forms a bridge between the transcription factors and RNA polymerase II. Although it has been reported that Med19 plays an important role in stabilizing the whole mediator complex, its biological importance in tongue cancer cell proliferation and migration has not been addressed. METHODS: By using MTT, BrdU incorporation, colony formation, flow cytometric, tumorigenesis and transwell assays, We tested the Med19 role on tongue cancer cell growth and migration...
2013: World Journal of Surgical Oncology
Keren Imberg-Kazdan, Susan Ha, Alex Greenfield, Christopher S Poultney, Richard Bonneau, Susan K Logan, Michael J Garabedian
The androgen receptor (AR) is a mediator of both androgen-dependent and castration-resistant prostate cancers. Identification of cellular factors affecting AR transcriptional activity could in principle yield new targets that reduce AR activity and combat prostate cancer, yet a comprehensive analysis of the genes required for AR-dependent transcriptional activity has not been determined. Using an unbiased genetic approach that takes advantage of the evolutionary conservation of AR signaling, we have conducted a genome-wide RNAi screen in Drosophila cells for genes required for AR transcriptional activity and applied the results to human prostate cancer cells...
April 2013: Genome Research
Hui Wen, Chen-chen Feng, Guan-xiong Ding, Dong-liang Meng, Qiang Ding, Zu-jun Fang, Guo-wei Xia, Gang Xu, Hao-wen Jiang
Bladder cancer (BCa) remained a major health problem. Med19 was related to tumor growth of BCa. Bone morphogenetic proteins (BMPs) were reported to be critical in bone metastasis of cancer. We therefore investigated the relations between Med19 and BMPs in BCa and their effect on bone metastasis of BCa. Bladder cancer cell lines were cultured and interfered with Med19 shRNA and control. Expressions of BMP-1, BMP-2, BMP-4, BMP-5, BMP-6, BMP-7, BMP-9, and BMP-15 were studied between 2 groups. Fifty-two BCa samples were included for immunohistochemical staining of Med19 and BMP-2...
June 2013: Annals of Diagnostic Pathology
Yingtao Liu, Xiang Tao, Lingling Fan, Luoqi Jia, Chao Gu, Youji Feng
Ovarian cancer causes more deaths than any other type of female reproductive cancer. The development of new therapeutic approaches is required due to the low survival rate using routine methods. The goal of this study was to investigate the effect of the gene silencing of mediator complex subunit 19 (MED19) on cell viability and tumor growth in ovarian cancer. Immunohistochemistry was used to characterize the expression of MED19 in human ovarian cancer tissues. Lentivirus-mediated RNAi was employed to downregulate endogenous MED19 expression in SKOV-3 and HEY ovarian cancer cells...
November 2012: Molecular Medicine Reports
Theresa Niederberger, Stefanie Etzold, Michael Lidschreiber, Kerstin C Maier, Dietmar E Martin, Holger Fröhlich, Patrick Cramer, Achim Tresch
The Mediator is a highly conserved, large multiprotein complex that is involved essentially in the regulation of eukaryotic mRNA transcription. It acts as a general transcription factor by integrating regulatory signals from gene-specific activators or repressors to the RNA Polymerase II. The internal network of interactions between Mediator subunits that conveys these signals is largely unknown. Here, we introduce MC EMiNEM, a novel method for the retrieval of functional dependencies between proteins that have pleiotropic effects on mRNA transcription...
2012: PLoS Computational Biology
Xiang-fu Ding, Guo-min Huang, Ying Shi, Jun-an Li, Xue-dong Fang
Med19 was a member of the Mediator complex which forms the bridge between transcriptional activators and RNA polymerase II. We aim to investigate the functional role of Med19 in the progression of human gastric carcinoma. The correlation between Med19 expression and clinicopathologic features in 60 gastric carcinoma specimens was analyzed by using immunohistochemistry. Recombinant lentivirus expressing Med19 short hairpin RNA (shRNA) was constructed and infected into human gastric carcinoma SGC7901 and MGC803 cells...
August 10, 2012: Gene
Monica Rienzo, Amelia Casamassimi, Concetta Schiano, Vincenzo Grimaldi, Teresa Infante, Claudio Napoli
Mediator (MED) is a fundamental component of the RNA polymerase II-mediated transcription machinery playing a pivotal role in the regulation of eukaryotic mRNA synthesis. Human MED complexes contain at least 30 distinct MED subunits. Our previous study, aimed to analyse MED complex during the pattern of endothelial progenitor cells (EPCs) differentiation, found an alternative transcript of MED30 subunit expressed only in circulating immature progenitor cells. Here, we report two novel transcripts of MED12 and MED19 subunits both generated by alternative splicing and displaying similar expression patterns, thereby indicating their involvement during endothelial cell differentiation...
August 2012: Biochimie
Selena B Kremer, Sunyoung Kim, Jeong Ok Jeon, Yara W Moustafa, Apeng Chen, Jing Zhao, David S Gross
Mediator is a modular multisubunit complex that functions as a critical coregulator of RNA polymerase II (Pol II) transcription. While it is well accepted that Mediator plays important roles in the assembly and function of the preinitiation complex (PIC), less is known of its potential roles in regulating downstream steps of the transcription cycle. Here we use a combination of genetic and molecular approaches to investigate Mediator regulation of Pol II elongation in the model eukaryote, Saccharomyces cerevisiae...
May 2012: Genetics
Xingang Cui, Danfeng Xu, Chao Lv, Fajun Qu, Jin He, Ming Chen, Yushan Liu, Yi Gao, Jianping Che, Yacheng Yao, Hongyu Yu
MED19 is a member of the Mediator that plays a key role in the activation and repression of signal transduction or the regulation of transcription in carcinomas. To tested the functional role of MED19 in human prostate cancer, we downregulated MED19 expression in prostate cancer cells (PC-3 and DU145) by lentivirus-mediated short hairpin (shRNA), and analyzed the effect of inhibition of MED19 on prostate cancer cell proliferation and tumorigenesis. The in vitro prostate cancer cell proliferation, colony formation, and in vivo tumor growth in nude mice xenografts was significantly reduced after the downregulation of MED19...
August 2011: BMB Reports
Xing-Hua Li, De-Ning Fang, Chai-Ming Zeng
Abnormal gene transcription plays an important role in oncogenesis. In cancer cells, the improper activation of specific genes is usually ascribed to aberrant transcription machinery including transcription factors, RNA polymerase II, and Mediator complex. This study reports on short hairpin RNA (shRNA)-mediated gene silencing of MED19, a subunit of Mediator complex, and its effect on the growth of pancreatic cancer cells. RNA interference was performed by lentivirus shRNA system to specifically knockdown MED19 expression in Aspc-1 and Panc-1 cells...
August 2011: Cancer Biotherapy & Radiopharmaceuticals
Tiejun Wang, Ling Hao, Yan Feng, Gang Wang, Darning Qin, Guishan Gu
MED 19 is a subunit of the mediator complex, which is a coactivator of RNA polymerase II and also interacts with the downstream coding region of many genes. However, the role of MED19 in osteosarcoma is unknown. In the present study, we applied lentivirus-mediated short hairpin RNA (shRNA)-triggered RNA interference to downregulate MED19 expression in human osteosarcoma SaOS-2 and U2OS cells. Knockdown of MED19 expression was confirmed by real-time PCR and Western blot. It was found that silencing of MED19 resulted in decreased cell viability, colony formation capacity, and DNA synthesis ability in both cells, as well as the G0/G1 phase cell cycle arrest...
2011: Oncology Research
Mei Sun, Rui Jiang, Jin-Dong Li, Shu-Li Luo, Hong-Wen Gao, Cheng-Yan Jin, Dong-Lei Shi, Chun-Guang Wang, Bin Wang, Xing-Yi Zhang
MED19 is a subunit of Mediator that is an essential component of RNA polymerase II-mediated transcription machinery. High expression levels of MED19 were examined in human lung adenocarcinoma tissues by immunohistochemical assay. MED19-specific short hairpin RNA (shRNA) expressing lentivirus was constructed and infected lung cancer cell line A549. MED19 mRNA and protein expression levels were downregulated in A549 cells as evidenced by real-time PCR and western blot assays. Importantly, MED19 inhibition resulted in impaired proliferation and colony formation, and induced accumulation of G1-phase cells and mitigated invasiveness of cells...
September 2011: Molecular and Cellular Biochemistry
Hu Zhang, Haowen Jiang, Wei Wang, Jian Gong, Limin Zhang, Zhongqing Chen, Qiang Ding
OBJECTIVES: The human Med19 gene encodes a critical subunit that stabilizes the whole mediator complex. To understand the role of Med19 in bladder cancer, we studied the effects of lentivirus-mediated suppression of Med19 expression on bladder cancer cells in vitro and in vivo. METHODS AND MATERIALS: In this study, immunohistochemical analysis was used to demonstrate the expression of Med19 in human bladder cancer. The lentivirus vectors containing a small hairpin RNA (shRNA) to target Med19 were constructed...
November 2012: Urologic Oncology
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