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https://www.readbyqxmd.com/read/28346086/a-small-molecule-microarray-approach-for-the-identification-of-e2-enzyme-inhibitors-in-ubiquitin-like-conjugation-pathways
#1
Katherine Zlotkowski, William M Hewitt, Ranu S Sinniah, Joseph E Tropea, Danielle Needle, George T Lountos, Joseph J Barchi, David S Waugh, John S Schneekloth
E2 enzymes in ubiquitin-like conjugation pathways are important, highly challenging pharmacological targets, and despite significant efforts, few noncovalent modulators have been discovered. Small-molecule microarray (SMM)-based screening was employed to identify an inhibitor of the "undruggable" small ubiquitin-like modifier (SUMO) E2 enzyme Ubc9. The inhibitor, a degradation product from a commercial screening collection, was chemically synthesized and evaluated in biochemical, mechanistic, and structure-activity relationship studies...
January 1, 2017: SLAS Discov
https://www.readbyqxmd.com/read/28342872/covalent-conjugation-of-the-equine-infectious-anemia-virus-gag-with-sumo
#2
Jinzhong Wang, Shuping Wen, Rui Zhao, Jing Qi, Zhao Liu, Weiwei Li, Jing An, Charles Wood, Ying Wang
The conjugation of small ubiquitin-like modifier (SUMO) to the target protein, namely, SUMOylation, is involved in the regulation of many important biological events including host-pathogen interaction. Some viruses have evolved to exploit the host SUMOylation machinery to modify their own protein. Retroviral Gag protein plays critical roles in the viral life cycle. The HIV-1 p6 and the Moloney murine leukemia virus CA have been reported to be conjugated with SUMO. In this study, we report for the first time, to our knowledge, the covalent conjugation of equine infectious anemia virus (EIAV) Gag with SUMO...
March 22, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28314856/ubc9-is-required-for-positive-selection-and-late-stage-maturation-of-thymocytes
#3
Aibo Wang, Xiao Ding, Maud Demarque, Xindong Liu, Deng Pan, Huawei Xin, Bo Zhong, Xiaohu Wang, Anne Dejean, Wei Jin, Chen Dong
SUMOylation is an important posttranslational modification that regulates protein function in diverse biological processes. However, its role in early T cell development has not been genetically studied. UBC9 is the only E2 enzyme for all SUMOylation. In this study, by selectively deleting Ubc9 gene in T cells, we have investigated the functional roles of SUMOylation in T cell development. Loss of Ubc9 results in a significant reduction of CD4 and CD8 single-positive lymphocytes in both thymus and periphery...
March 17, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28285006/gata5-sumoylation-is-indispensable-for-zebrafish-cardiac-development
#4
Bin Wen, Hao Yuan, Xiaohui Liu, Haihong Wang, Saijuan Chen, Zhu Chen, Hugues de The, Jun Zhou, Jun Zhu
BACKGROUND: SUMOylation is a critical regulatory protein modification in eukaryotic cells and plays a pivotal role in cardiac development and disease. Several cardiac transcription factors are modified by SUMO, but little is known about the impact of SUMOylation on their function during cardiac development. METHODS: We used a zebrafish model to address the impact of SUMOylation on GATA5, an essential transcription factor in zebrafish cardiac development. GATA5 SUMOylation was probed by western blot, the subcellular localization and transcriptional activity of GATA5 mutants were examined by immunostaining and luciferase reporter assay...
March 8, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28253371/autophagy-regulates-ubc9-levels-during-viral-mediated-tumorigenesis
#5
Domenico Mattoscio, Chiara Casadio, Claudia Miccolo, Fausto Maffini, Andrea Raimondi, Carlo Tacchetti, Tarik Gheit, Marta Tagliabue, Viviana E Galimberti, Francesca De Lorenzi, Michael Pawlita, Fausto Chiesa, Mohssen Ansarin, Massimo Tommasino, Susanna Chiocca
UBC9, the sole E2-conjugating enzyme required for SUMOylation, is a key regulator of essential cellular functions and, as such, is frequently altered in cancers. Along these lines, we recently reported that its expression gradually increases during early stages of human papillomavirus (HPV)-mediated cervical lesions transformation. However, a better understanding of how UBC9 is exploited by transforming viral oncoproteins is still needed. In the present study, we show that in human samples HPV drives UBC9 up-regulation also in very early steps of head and neck tumorigenesis, pointing to the important role for UBC9 in the HPV-mediated carcinogenic program...
March 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28247205/the-mechanism-of-jurkat-cells-apoptosis-induced-by-aggregatibacter-actinomycetemcomitans-cytolethal-distending-toxin
#6
Hui-Ping Chen, Lu Li, Xu Chen, Mi-Fang Yang, Yu Ye, Xiao-Qian Wang, Yan Xu
Cytolethal distending toxin (CDT) which is produced by Aggregatibacter actinomycetemcomitans causes apoptosis in lymphocytes. But the specific mechanism is not clear. The aim of our research was to investigate the effect and mechanism during this process. The wild-type CdtA, CdtB, CdtC (CdtA(W), CdtB(W), CdtC(W)) and mutant CdtB (CdtB(M)) were expressed and purified respectively and the purity of each subunit was examined by BandScan software. And the type I deoxyribonuclease and PI-3,4,5-triphosphate (PI-3,4,5-P3, PIP3) phosphatase activity were detected by DNA agarose gel electrophoresis and enzyme-linked immunosorbent assay respectively...
February 28, 2017: Apoptosis: An International Journal on Programmed Cell Death
https://www.readbyqxmd.com/read/28240261/nur77-suppresses-hepatocellular-carcinoma-via-switching-glucose-metabolism-toward-gluconeogenesis-through-attenuating-phosphoenolpyruvate-carboxykinase-sumoylation
#7
Xue-Li Bian, Hang-Zi Chen, Peng-Bo Yang, Ying-Ping Li, Fen-Na Zhang, Jia-Yuan Zhang, Wei-Jia Wang, Wen-Xiu Zhao, Sheng Zhang, Qi-Tao Chen, Yu Zheng, Xiao-Yu Sun, Xiao-Min Wang, Kun-Yi Chien, Qiao Wu
Gluconeogenesis, an essential metabolic process for hepatocytes, is downregulated in hepatocellular carcinoma (HCC). Here we show that the nuclear receptor Nur77 is a tumour suppressor for HCC that regulates gluconeogenesis. Low Nur77 expression in clinical HCC samples correlates with poor prognosis, and a Nur77 deficiency in mice promotes HCC development. Nur77 interacts with phosphoenolpyruvate carboxykinase (PEPCK1), the rate-limiting enzyme in gluconeogenesis, to increase gluconeogenesis and suppress glycolysis, resulting in ATP depletion and cell growth arrest...
February 27, 2017: Nature Communications
https://www.readbyqxmd.com/read/28197924/sumoylation-as-an-integral-mechanism-in-bacterial-infection-and-disease-progression
#8
Chittur V Srikanth, Smriti Verma
Post translational modification pathways regulate fundamental processes of cells and thus govern vital functions. Among these, particularly the modification with Small Ubiquitin-like Modifiers (SUMO) is being recognized as a pathway crucial for cell homeostasis and health. Understandably, bacterial pathogens intervene with the SUMO pathway of the host for ensuring successful infection. Among the bacterial pathogens known to target host sumoylation varied points of intervention are utilized. Majority of them including Salmonella Typhimurium, Shigella flexneri and Listeria monocytogenes target the E2 conjugating enzyme Ubc9...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28195188/sumo-regulates-the-activity-of-smoothened-and-costal-2-in-drosophila-hedgehog-signaling
#9
Jie Zhang, Yajuan Liu, Kai Jiang, Jianhang Jia
In Hedgehog (Hh) signaling, the GPCR-family protein Smoothened (Smo) acts as a signal transducer that is regulated by phosphorylation and ubiquitination, which ultimately change the cell surface accumulation of Smo. However, it is not clear whether Smo is regulated by other post-translational modifications, such as sumoylation. Here, we demonstrate that knockdown of the small ubiquitin-related modifier (SUMO) pathway components Ubc9 (a SUMO-conjugating enzyme E2), PIAS (a SUMO-protein ligase E3), and Smt3 (the SUMO isoform in Drosophila) by RNAi prevents Smo accumulation and alters Smo activity in the wing...
February 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28150190/estrogen-modulates-ubc9-expression-and-synaptic-redistribution-in-the-brain-of-app-ps1-mice-and-cortical-neurons
#10
Yu-Jie Lai, Lu Liu, Xiao-Tong Hu, Ling He, Guo-Jun Chen
Estrogen exerts multiple actions in the brain and is an important neuroprotective factor in a number of neuronal disorders. However, the underlying mechanism remains unknown. Studies demonstrate that ubiquitin-conjugating enzyme 9 (ubc9) has an integral role in synaptic plasticity and may contribute to the pathology of neuronal disorders. We aimed to investigate the effects of estrogen on ubc9 and in the Alzheimer's disease brain. Ubc9 protein and mRNA were significantly increased in the cortex and hippocampus of APP/PS1 mice with enhanced SUMOylation...
February 1, 2017: Journal of Molecular Neuroscience: MN
https://www.readbyqxmd.com/read/28143738/manipulating-pml-sumoylation-via-silencing-ubc9-and-rnf4-regulates-cardiac-fibrosis
#11
Yu Liu, Dan Zhao, Fang Qiu, Ling-Ling Zhang, Shang-Kun Liu, Yuan-Yuan Li, Mei-Tong Liu, Di Wu, Jia-Xin Wang, Xiao-Qing Ding, Yan-Xin Liu, Chang-Jiang Dong, Xiao-Qi Shao, Bao-Feng Yang, Wen-Feng Chu
The promyelocytic leukemia protein (PML) is essential in the assembly of dynamic subnuclear structures called PML nuclear bodies (PML-NBs), which are involved in regulating diverse cellular functions. However, the possibility of PML being involved in cardiac disease has not been examined. In mice undergoing transverse aortic constriction (TAC) and arsenic trioxide (ATO) injection, transforming growth factor β1 (TGF-β1) was upregulated along with dynamic alteration of PML SUMOylation. In cultured neonatal mouse cardiac fibroblasts (NMCFs), ATO, angiotensin II (Ang II), and fetal bovine serum (FBS) significantly triggered PML SUMOylation and the assembly of PML-NBs...
March 1, 2017: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/28132806/rifampicin-pre-treatment-inhibits-the-toxicity-of-rotenone-induced-pc12-cells-by-enhancing-sumoylation-modification-of-%C3%AE-synuclein
#12
D Lin, X Jing, Y Chen, Y Liang, M Lei, S Peng, T Zhou, D Zheng, Z Zeng, X Wu, L Yang, S Xiao, J Liu, E Tao
Our previous research revealed that rifampicin could protect PC12 (pheochromocytoma 12) cells from rotenone-induced cytotoxicity by reversing the aggregation of α-synuclein. Furthermore, increasing evidence indicated that the misfolded α-synuclein with SUMOylation, an important protein posttranslational modification, was easier to solubilize and was less toxic. Here, we investigated whether rifampicin could stabilize α-synuclein and prevent rotenone-induced PC12 cells from undergoing apoptosis by enhancing SUMOylation of α-synuclein...
March 25, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28078755/modulation-of-host-cell-sumoylation-facilitates-efficient-development-of-plasmodium-berghei-and-toxoplasma-gondii
#13
Mulaka Maruthi, Dipti Singh, Segireddy Rameswara Reddy, Babu S Mastan, Satish Mishra, Kota Arun Kumar
SUMOylation is a reversible post translational modification of proteins that regulates protein stabilization, nucleocytoplasmic transport, and protein-protein interactions. Several viruses and bacteria modulate host SUMOylation machinery for efficient infection. Plasmodium sporozoites are infective forms of malaria parasite that invade mammalian hepatocytes and transforms into exoerythrocytic forms (EEFs). Here, we show that during EEF development, the distribution of SUMOylated proteins in host cell nuclei was significantly reduced and expression of the SUMOylation enzymes was downregulated...
January 12, 2017: Cellular Microbiology
https://www.readbyqxmd.com/read/27993970/identification-of-a-multifunctional-docking-site-on-the-catalytic-unit-of-phosphodiesterase-4-pde4-that-is-utilised-by-multiple-interaction-partners
#14
Kirsty F Houslay, Frank Christian, Ruth MacLeod, David R Adams, Miles D Houslay, George S Baillie
Cyclic AMP (cAMP)-specific phosphodiesterase-4 (PDE4) enzymes underpin compartmentalised cAMP signalling by localising to distinct signalling complexes. PDE4 long isoforms can be phosphorylated by mitogen-activated protein kinase-activated protein kinase 2 (MK2), which attenuates activation of such enzymes through their phosphorylation by protein kinase A. Here we show that MK2 interacts directly with PDE4 long isoforms and define the sites of interaction. One is a unique site that locates within the regulatory upstream conserved region 1 (UCR1) domain and contains a core Phe141, Leu142 and Tyr143 (FLY) cluster (PDE4A5 numbering)...
February 15, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/27889610/cacul1-cac1-attenuates-p53-activity-through-pml-post-translational-modification
#15
Tomomi Fukuda, Yu Kigoshi-Tansho, Takao Naganuma, Akira Kazaana, Tomomi Okajima, Fuminori Tsuruta, Tomoki Chiba
Promyelocytic leukaemia (PML) is a tumor suppressor protein covalently conjugated with SUMO family proteins, leading to the formation of PML nuclear bodies (NBs). PML-NBs provide a platform for efficient posttranslational modification of targets and protein-protein interaction, contributing to the adjustment of gene expression and chromatin integrity. Although PML SUMOylation is thought to play important roles in diverse cellular functions, the control mechanisms of adequate modification levels have remained unsolved...
January 22, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27878232/involvement-of-bleomycin-hydrolase-and-poly-adp-ribose-polymerase-1-in-ubc9-mediated-resistance-to-chemotherapy-agents
#16
Yang Chen, Huixian Zhang, Qiyang He
Ubiquitin-conjugating protein 9 (Ubc9), the sole enzyme for sumoylation, plays critical roles in many physiological functions, such as DNA damage repair and genome integrity. Its overexpression led to poor prognosis and drug resistance in tumor chemotherapy. However, the underlying mechanism by which Ubc9 promotes tumor progress and influences the susceptibility to antitumor agents remains elusive. In this study, we used nine antitumor agents with distinct actions to explore Ubc9-mediated resistance in human breast carcinoma MCF-7 cells...
January 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/27806335/orthoreovirus-outer-fiber-proteins-are-substrates-for-sumo-conjugating-enzyme-ubc9
#17
Fei Yu, Hao Wang, Longlong Wang, Liqun Lu
Reoviruses are potential anticancer agents due to their ability to induce cell death in tumor cells. Grass carp reovirus (GCRV) is one of the best characterized models on reovirus pathogenesis in vitro. However, there is little known about how SUMOylation affects reovirus pathogenesis. The SUMO conjugating enzyme 9 (Ubc9) determines the targets of SUMOylation. Here, the protein interactions between reovirus outer fiber proteins, specifically GCRV-104 VP55, and Ubc9 were probed using a yeast two-hybrid system...
October 28, 2016: Oncotarget
https://www.readbyqxmd.com/read/27758866/mitochondrial-activity-in-human-white-adipocytes-is-regulated-by-the-ubiquitin-carrier-protein-9-microrna-30a-axis
#18
Eun Hee Koh, Yong Chen, David A Bader, Mark P Hamilton, Bin He, Brian York, Shingo Kajimura, Sean E McGuire, Sean M Hartig
The acquisition of beige adipocyte features by white fat cells corresponds to protection against obesity-induced metabolic diseases in humans and animal models of type 2 diabetes. In adipose tissue, expression of the E2 small ubiquitin-like modifier ligase ubiquitin carrier protein 9 (Ubc9) is positively correlated with markers of insulin resistance and corresponds with impaired browning of human white adipocytes. However, the molecular regulation of Ubc9 expression in adipocytes and other cells remains unclear...
November 18, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27658615/sumoylation-of-ikb-attenuates-nf-kb-induced-nitrosative-stress-at-rostral-ventrolateral-medulla-and-cardiovascular-depression-in-experimental-brain-death
#19
Ching-Yi Tsai, Faith C H Li, Carol H Y Wu, Alice Y W Chang, Samuel H H Chan
BACKGROUND: Small ubiquitin-related modifier (SUMO) is a group of proteins that participates in post-translational modifications. One known SUMO target is the transcription factor nuclear factor-kB (NF-kB) that plays a pivotal role in many disease processes; sumoylation inactivates NF-kB by conjugation with inhibitors of NF-kB (IkB). Our laboratory demonstrated previously that transcriptional upregulation of nitric oxide synthase II (NOS II) by NF-kB, leading to nitrosative stress by the formation of peroxynitrite in the rostral ventrolateral medulla (RVLM), underpins the defunct brain stem cardiovascular regulation that precedes brain death...
2016: Journal of Biomedical Science
https://www.readbyqxmd.com/read/27631796/reconstitution-of-the-recombinant-ranbp2-sumo-e3-ligase-complex
#20
Tobias Ritterhoff, Hrishikesh Das, Yuqing Hao, Volkan Sakin, Annette Flotho, Andreas Werner, Frauke Melchior
One of the few proteins that have SUMO E3 ligase activity is the 358 kDa nucleoporin RanBP2 (Nup358). While small fragments of RanBP2 can stimulate SUMOylation in vitro, the physiologically relevant E3 ligase is a stable multi-subunit complex comprised of RanBP2, SUMOylated RanGAP1, and Ubc9. Here, we provide a detailed protocol to in vitro reconstitute the RanBP2 SUMO E3 ligase complex. With the exception of RanBP2, reconstitution involves untagged full-length proteins. We describe the bacterial expression and purification of all complex components, namely an 86 kDa His-tagged RanBP2 fragment, the SUMO E2-conjugating enzyme Ubc9, RanGAP1, and SUMO1, and we provide a protocol for quantitative SUMOylation of RanGAP1...
2016: Methods in Molecular Biology
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