DOCSLIB.ORG

USP8 (D18F6) Rabbit Mab - 20ºC Store at Support: 877-678-TECH (8324) [email protected]

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
USP8 (D18F6) Rabbit Mab - 20ºC Store at Support: 877-678-TECH (8324) Info@Cellsignal.Com www.cellsignal.com 20ºC USP8 (D18F6) Rabbit mAb - Store at Support: 877-678-TECH (8324) [email protected] 100 µl (10 western blots) Orders: 877-616-CELL (2355) [email protected] Entrez-Gene ID #9101 #11832 rev. 06/09/14 UniProt ID #P40818 For Research Use Only. Not For Use In Diagnostic Procedures. Applications Species Cross-Reactivity* Molecular Wt. Isotype Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% W, IP H, Mk 140 kDa Rabbit IgG** Endogenous sodium azide. Store at –20°C. Do not aliquot the antibody. *Species cross-reactivity is determined by western blot. ** Anti-rabbit secondary antibodies must be used to Background: Ubiquitinating enzymes (UBEs) catalyze protein kDa detect this antibody. ubiquitination, a reversible process countered by deubiquiti- nating enzyme (DUB) action (1,2). Five DUB subfamilies are 200 Recommended Antibody Dilutions: recognized, including the USP, UCH, OTU, MJD, and JAMM Western blotting 1:1000 enzymes. The deubiquitinating enzyme ubiquitin-specific USP8 (Myc/DDK-tagged) Immunoprecipitation 1:100 protease 8 (USP8/UBPy) is a cysteine protease belonging to the 140 USP8 (Endogenous) Background References: USP/UBP subfamily. Research studies have shown that USP8 is an essential growth-regulated enzyme indespensible for cell (1) Nijman, S.M. et al. (2005) Cell 123, 773-86. 100 proliferation and survival (3,4). Indeed, conditional knock-out of (2) Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613. murine USP8 was shown to promote a dramatic loss in expres- (3) Naviglio, S. et al. (1998) EMBO J 17, 3241-50. sion of receptor tyrosine kinases, including EGFR, ErbB3, and 80 c-Met (4). In agreement with these findings, USP8 inactivation – + hUSP8-Myc/DDK (4) Niendorf, S. et al. (2007) Mol Cell Biol 27, 5029-39. leads to enhanced ubiquitination of ligand-activated EGFR (5,6). Western blot analysis of extracts from 293T cells, either mock (5) Alwan, H.A. and van Leeuwen, J.E. (2007) J Biol Chem Furthermore, phosphorylation of USP8 at Ser680 results in its 282, 1658-69. binding of 14-3-3, catalytic inactivation, and reduced EGFR transfected (-) or transfected with a construct expressing Myc/DDK- tagged full-length human USP8 (hUSP8-Myc/DDK +), using USP8 deubiquitination (7). It appears as though USP8, in conjunction (6) Mizuno, E. et al. (2005) Mol Biol Cell 16, 5163-74. (D18F6) Rabbit mAb. with components of the ESCRT-0 complex, plays an integral (7) Mizuno, E. et al. (2007) Exp Cell Res 313, 3624-34. role in the early endosomal sorting machinery that functions to (8) Row, P.E. et al. (2006) J Biol Chem 281, 12618-24. protect EGFR from lysosomal degradation (8,9). (9) Berlin, I. et al. (2010) J Biol Chem 285, 34909-21. Specificity/Sensitivity: USP8 (D18F6) Rabbit mAb recog- nizes endogenous levels of total USP8 protein. Source/Purification: Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu614 of human USP8 protein. kDa 293T MCF7 U-2 OS NCI-H460DLD-1 A549 U251 KARPAS-299A-431 A-375 COS-7 Western blot analysis of extracts from various cell lines 200 using USP8 (D18F6) Rabbit mAb. 140 USP8 100 80 Tween® is a registered trademark of ICI Americas, Inc. Thank you for your recent purchase. If you would like to provide a review visit www.cellsignal.com/comments. IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight. © 2014 Cell Signaling Technology, Inc. Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc. Applications: W—Western IP—Immunoprecipitation IHC—Immunohistochemistry ChIP—Chromatin Immunoprecipitation IF—Immunofluorescence F—Flow cytometry E-P—ELISA-Peptide Species Cross-Reactivity: H—human M—mouse R—rat Hm—hamster Mk—monkey Mi—mink C—chicken Dm—D. melanogaster X—Xenopus Z—zebrafish B—bovine Dg—dog Pg—pig Sc—S. cerevisiae Ce—C. elegans Hr—Horse All—all species expected Species enclosed in parentheses are predicted to react based on 100% homology..
Load more

Recommended publications
  • USP8 Antibody A
    Revision 1 C 0 2 - t USP8 Antibody a e r o t S Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) 8 2 Web: [email protected] 7 www.cellsignal.com 8 # 3 Trask Lane Danvers Massachusetts 01923 USA For Research Use Only. Not For Use In Diagnostic Procedures. Applications: Reactivity: Sensitivity: MW (kDa): Source: UniProt ID: Entrez-Gene Id: WB, IP H Mk Endogenous 130 Rabbit P40818 9101 Product Usage Information 6. Mizuno, E. et al. (2005) Mol Biol Cell 16, 5163-74. 7. Mizuno, E. et al. (2007) Exp Cell Res 313, 3624-34. Application Dilution 8. Row, P.E. et al. (2006) J Biol Chem 281, 12618-24. 9. Berlin, I. et al. (2010) J Biol Chem 285, 34909-21. Western Blotting 1:1000 Immunoprecipitation 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody. Specificity / Sensitivity USP8 Antibody recognizes endogenous levels of total USP8 protein. Species Reactivity: Human, Monkey Source / Purification Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro320 of human USP8 protein. Antibodies are purified by protein A and peptide affinity chromatography. Background Ubiquitinating enzymes (UBEs) catalyze protein ubiquitination, a reversible process countered by deubiquitinating enzyme (DUB) action (1,2). Five DUB subfamilies are recognized, including the USP, UCH, OTU, MJD, and JAMM enzymes. The deubiquitinating enzyme ubiquitin-specific protease 8 (USP8/UBPy) is a cysteine protease belonging to the USP/UBP subfamily.
    [Show full text]
  • Clec16a, Nrdp1, and USP8 Form a Ubiquitin-Dependent
    Page 1 of 46 Diabetes Clec16a, Nrdp1, and USP8 form a ubiquitin-dependent tripartite complex that regulates beta cell mitophagy Gemma Pearson1; Biaoxin Chai1; Tracy Vozheiko1; Xueying Liu1; Malathi Kandarpa2; 3 1,4* Robert C. Piper ; and Scott A. Soleimanpour 1Division of Metabolism, Endocrinology & Diabetes, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48105, USA. 2Division of Hematology and Oncology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48105, USA. 3Department of Molecular Physiology and Biophysics, University of Iowa Carver College of Medicine, Iowa City, IA 52242, USA 4Veterans Affairs Ann Arbor Health Care System, Ann Arbor, MI 48105, USA. *Corresponding Author Scott A. Soleimanpour, M.D., 1000 Wall Street, 5317 Brehm Tower, Ann Arbor, MI 48105. Phone: (734) 763-0528 Fax: (734) 232-8162 E-mail: [email protected] Running Title: β-cell mitophagy requires ubiquitin signaling Word Count: 4448 Figures: 7 Diabetes Publish Ahead of Print, published online November 27, 2017 Diabetes Page 2 of 46 ABSTRACT Mitophagy is a cellular quality control pathway, which is essential to eliminate unhealthy mitochondria. While mitophagy is critical to pancreatic β-cell function, the post-translational signals governing β-cell mitochondrial turnover are unknown. Here we report that ubiquitination is essential for the assembly of a mitophagy regulatory complex, comprised of the E3 ligase Nrdp1, the deubiquitinase enzyme USP8, and Clec16a, a mediator of β-cell mitophagy with unclear function. We discover that the diabetes gene Clec16a encodes an E3 ligase, which promotes non-degradative ubiquitin conjugates to direct its mitophagy effectors and stabilize the Clec16a-Nrdp1-USP8 complex.
    [Show full text]
  • Hereditary Spastic Paraplegia: from Genes, Cells and Networks to Novel Pathways for Drug Discovery
    brain sciences Review Hereditary Spastic Paraplegia: From Genes, Cells and Networks to Novel Pathways for Drug Discovery Alan Mackay-Sim Griffith Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia; a.mackay-sim@griffith.edu.au Abstract: Hereditary spastic paraplegia (HSP) is a diverse group of Mendelian genetic disorders affect- ing the upper motor neurons, specifically degeneration of their distal axons in the corticospinal tract. Currently, there are 80 genes or genomic loci (genomic regions for which the causative gene has not been identified) associated with HSP diagnosis. HSP is therefore genetically very heterogeneous. Finding treatments for the HSPs is a daunting task: a rare disease made rarer by so many causative genes and many potential mutations in those genes in individual patients. Personalized medicine through genetic correction may be possible, but impractical as a generalized treatment strategy. The ideal treatments would be small molecules that are effective for people with different causative mutations. This requires identification of disease-associated cell dysfunctions shared across geno- types despite the large number of HSP genes that suggest a wide diversity of molecular and cellular mechanisms. This review highlights the shared dysfunctional phenotypes in patient-derived cells from patients with different causative mutations and uses bioinformatic analyses of the HSP genes to identify novel cell functions as potential targets for future drug treatments for multiple genotypes. Keywords: neurodegeneration; motor neuron disease; spastic paraplegia; endoplasmic reticulum; Citation: Mackay-Sim, A. Hereditary protein-protein interaction network Spastic Paraplegia: From Genes, Cells and Networks to Novel Pathways for Drug Discovery. Brain Sci. 2021, 11, 403.
    [Show full text]
  • Differential Microrna Expression in USP8-Mutated and Wild-Type Corticotroph Pituitary Tumors Reflect the Difference in Protein Ubiquitination Processes
    Journal of Clinical Medicine Article Differential microRNA Expression in USP8-Mutated and Wild-Type Corticotroph Pituitary Tumors Reflect the Difference in Protein Ubiquitination Processes Mateusz Bujko 1 , Paulina Kober 1, Joanna Boresowicz 1, Natalia Rusetska 1, Natalia Zeber-Lubecka 2, Agnieszka Paziewska 3, Monika Pekul 4, Grzegorz Zielinski 5, Andrzej Styk 5, Jacek Kunicki 6, Jerzy Ostrowski 2,7, Janusz A Siedlecki 1 and Maria Maksymowicz 4,* 1 Department of Molecular and Translational Oncology, Maria Sklodowska-Curie National Research Institute of Oncology, 02-781 Warsaw, Poland; [email protected] (M.B.); [email protected] (P.K.); [email protected] (J.B.); [email protected] (N.R.); [email protected] (J.A.S.) 2 Department of Gastroenterology, Hepatology and Clinical Oncology, Medical Centre for Postgraduate Education, 01-813 Warsaw, Poland; [email protected] (N.Z.-L.); [email protected] (J.O.) 3 Department of Neuroendocrinology, Centre of Postgraduate Medical Education, 01-813 Warsaw, Poland; [email protected] 4 Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Institute-Oncology Center, 02-781 Warsaw, Poland; [email protected] 5 Department of Neurosurgery, Military Institute of Medicine, 04-141 Warsaw, Poland; [email protected] (G.Z.); [email protected] (A.S.) 6 Department of Neurosurgery, Maria Sklodowska-Curie Institute-Oncology Center, 02-781 Warsaw, Poland; [email protected] Citation: Bujko, M.; Kober, P.; 7 Department of Genetics, Maria Sklodowska-Curie Institute-Oncology Center, 02-781 Warsaw, Poland Boresowicz, J.; Rusetska, N.; * Correspondence: [email protected] or [email protected] Zeber-Lubecka, N.; Paziewska, A.; Pekul, M.; Zielinski, G.; Styk, A.; Abstract: Background: USP8 mutations are the most common driver changes in corticotroph pituitary Kunicki, J.; et al.
    [Show full text]
  • Identification of a Novel RASD1 Somatic Mutation in a USP8-Mutated Corticotroph Adenoma
    Downloaded from molecularcasestudies.cshlp.org on September 27, 2021 - Published by Cold Spring Harbor Laboratory Press TITLE PAGE Title: Identification of a novel RASD1 somatic mutation in a USP8-mutated corticotroph adenoma. Running title: Novel RASD1 mutation in a corticotroph adenoma. Article Type: Research Report Authors and affiliations: Andrew V. Uzilov 1 Khadeen C. Cheesman* 2 Marc Y. Fink* 1 Leah C. Newman 1 Chetanya Pandya 1 Yelena Lalazar 2 Marco Hefti 3 Mary Fowkes 3 Gintaras Deikus 1 Chun Yee Lau 1 Aye S. Moe 1 Yayoi Kinoshita 3 Yumi Kasai 1 4 Micol Zweig 1 Arpeta Gupta 2 Daniela Starcevic 1 Milind Mahajan 1 Eric E. Schadt 1 Kalmon D. Post 5 Michael J. Donovan 3 Robert Sebra 1 Rong Chen# 1 Eliza B. Geer# 2,5,6 * Contributed equally to this work. # corresponding author, [email protected], [email protected] 1) Department of Genetics and Genomic Sciences and Icahn Institute for Genomics and Multiscale Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA 2) Division of Endocrinology, Diabetes, and Bone Disease, Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA Downloaded from molecularcasestudies.cshlp.org on September 27, 2021 - Published by Cold Spring Harbor Laboratory Press 3) Department of Pathology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA 4) Current affiliation: New York Genome Center, New York, NY 10013, USA 5) Department of Neurosurgery, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA 6) Multidisciplinary Pituitary and Skull Base Tumor Center, Memorial Sloan Kettering, 1275 York Avenue, New York, NY 10065, USA ABSTRACT Cushing’s Disease is caused by pituitary corticotroph adenomas that secrete excess adrenocorticotropic hormone.
    [Show full text]
  • Update on the Genetics of Spastic Paraplegias Maxime Boutry, Sara Morais, Giovanni Stevanin
    Update on the genetics of spastic paraplegias Maxime Boutry, Sara Morais, Giovanni Stevanin To cite this version: Maxime Boutry, Sara Morais, Giovanni Stevanin. Update on the genetics of spastic paraple- gias. Current Neurology and Neuroscience Reports, Current Medicine Group, 2019, 19 (4), pp.18. 10.1007/s11910-019-0930-2. hal-02272013 HAL Id: hal-02272013 https://hal.sorbonne-universite.fr/hal-02272013 Submitted on 27 Aug 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Update on the genetics of spastic paraplegias Maxime Boutry1,2*, Sara Morais1,2,3, Giovanni Stevanin1,2 1 Institut du Cerveau et de la Moelle épinière, Sorbonne Université UMR_S1127, INSERM Unit 1127, CNRS UMR7225, 75013, Paris, France 2 Ecole Pratique des Hautes Etudes (EPHE), Paris Sciences Lettres (PSL) Research University, Neurogenetics team, 75013, Paris, France 3 UnIGENe, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal *Current affiliation: Cell Biology Program, Hospital for Sick Children, Toronto, Ontario, Canada Corresponding author: Giovanni Stevanin [email protected] ORCID 0000-0001-9368-8657 Abstract Purpose of review Hereditary spastic paraplegias are a genetically heterogeneous group of neurological disorders.
    [Show full text]
  • Systematic Elucidation of Neuron-Astrocyte Interaction in Models of Amyotrophic Lateral Sclerosis Using Multi-Modal Integrated Bioinformatics Workflow
    ARTICLE https://doi.org/10.1038/s41467-020-19177-y OPEN Systematic elucidation of neuron-astrocyte interaction in models of amyotrophic lateral sclerosis using multi-modal integrated bioinformatics workflow Vartika Mishra et al.# 1234567890():,; Cell-to-cell communications are critical determinants of pathophysiological phenotypes, but methodologies for their systematic elucidation are lacking. Herein, we propose an approach for the Systematic Elucidation and Assessment of Regulatory Cell-to-cell Interaction Net- works (SEARCHIN) to identify ligand-mediated interactions between distinct cellular com- partments. To test this approach, we selected a model of amyotrophic lateral sclerosis (ALS), in which astrocytes expressing mutant superoxide dismutase-1 (mutSOD1) kill wild-type motor neurons (MNs) by an unknown mechanism. Our integrative analysis that combines proteomics and regulatory network analysis infers the interaction between astrocyte-released amyloid precursor protein (APP) and death receptor-6 (DR6) on MNs as the top predicted ligand-receptor pair. The inferred deleterious role of APP and DR6 is confirmed in vitro in models of ALS. Moreover, the DR6 knockdown in MNs of transgenic mutSOD1 mice attenuates the ALS-like phenotype. Our results support the usefulness of integrative, systems biology approach to gain insights into complex neurobiological disease processes as in ALS and posit that the proposed methodology is not restricted to this biological context and could be used in a variety of other non-cell-autonomous communication
    [Show full text]
  • MAGEL2 Regulates Leptin Receptor Internalization Through Ubiquitination Pathways Involving USP8 and RNF41
    MAGEL2 regulates leptin receptor internalization through ubiquitination pathways involving USP8 and RNF41 by Tishani Methsala Wijesuriya A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science Medical Sciences – Medical Genetics University of Alberta ©Tishani Methsala Wijesuriya, 2017 Abstract Children with Prader-Willi syndrome have neonatal feeding difficulties, developmental delay and excessive appetite. Loss of MAGEL2 alone causes a related neurodevelopmental disorder (Schaaf- Yang syndrome) and may contribute to obesity in children with Prader-Willi syndrome who lack MAGEL2 and other genes. MAGEL2 is essential in neurons that sense levels of the adipose tissue- derived hormone leptin. The MAGEL2 protein is important for recycling or degradation of proteins in the brain and interacts with and modifies the activity of E3 ubiquitin ligases. RNF41 is a E3 ubiquitin ligase that associates with a ubiquitin-specific protease (USP8). Together with USP8, RNF41 regulates the recycling of the leptin receptor by targeting it either for degradation or for recycling to the cell membrane. I hypothesized that MAGEL2 regulates the interaction between RNF41 and USP8, and that loss of this regulation could impair leptin response pathways in the brain in children with PWS. Human U2OS cells were transfected with recombinant constructs encoding epitope tagged versions of MAGEL2 and wild type and mutant forms of RNF41. Immunofluorescence was used to visualize the co-localization of different forms of RNF41 with MAGEL2 in intracellular compartments. I expressed recombinant MAGEL2, RNF41 and USP8 in combinations in human U2OS cells and examined the relative abundance of each protein in the presence or absence of the other components of the complex.
    [Show full text]
  • Reciprocal Cross-Regulation Between RNF41 and USP8 Controls Cytokine
    3770 Research Article Reciprocal cross-regulation between RNF41 and USP8 controls cytokine receptor sorting and processing Leentje De Ceuninck1,2,*, Joris Wauman1,2,*, Delphine Masschaele1,2, Frank Peelman1,2 and Jan Tavernier1,2,` 1Department of Medical Protein Research, Flanders Interuniversity Institute for Biotechnology (VIB), Albert Baertsoenkaai 3, B-9000 Ghent, Belgium 2Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University, Albert Baertsoenkaai 3, B-9000 Ghent, Belgium *These authors contributed equally to this work `Author for correspondence ([email protected]) Accepted 15 May 2013 Journal of Cell Science 126, 3770–3781 ß 2013. Published by The Company of Biologists Ltd doi: 10.1242/jcs.131250 Summary The mechanisms controlling the steady-state cell surface levels of cytokine receptors, and consequently the cellular response to cytokines, remain poorly understood. The number of surface-exposed receptors is a dynamic balance of de novo synthesis, transport to the plasma membrane, internalization, recycling, degradation and ectodomain shedding. We previously reported that the E3 ubiquitin ligase RING finger protein 41 (RNF41) inhibits basal lysosomal degradation and enhances ectodomain shedding of JAK2-associated cytokine receptors. Ubiquitin-specific protease 8 (USP8), an RNF41-interacting deubiquitylating enzyme (DUB) stabilizes RNF41 and is involved in trafficking of various transmembrane proteins. The present study identifies USP8 as a substrate of RNF41 and reveals that loss of USP8 explains the aforementioned RNF41 effects. RNF41 redistributes and ubiquitylates USP8, and reduces USP8 levels. In addition, USP8 knockdown functionally matches the effects of RNF41 ectopic expression on the model leptin and leukemia inhibitory factor (LIF) receptors. Moreover, RNF41 indirectly destabilizes the ESCRT-0 complex through suppression of USP8.
    [Show full text]
  • Hereditary Spastic Paraplegias: Clinical Spectrum in Sudan, Further Deciphering of the Molecular Bases of Autosomal Recessive Forms and New Genes Emerging
    Hereditary spastic paraplegias : clinical spectrum in Sudan, further deciphering of the molecular bases of autosomal recessive forms and new genes emerging Liena Elbaghir Omer Elsayed To cite this version: Liena Elbaghir Omer Elsayed. Hereditary spastic paraplegias : clinical spectrum in Sudan, further deciphering of the molecular bases of autosomal recessive forms and new genes emerging. Neurons and Cognition [q-bio.NC]. Université Pierre et Marie Curie - Paris VI; University of Khartoum, 2016. English. NNT : 2016PA066056. tel-01438739 HAL Id: tel-01438739 https://tel.archives-ouvertes.fr/tel-01438739 Submitted on 18 Jan 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Université Pierre et Marie Curie University of Khartoum Cerveau-Cognition-Comportement (ED3C) Institut du Cerveau et de la Moelle Epinière / Equipe Bases Moléculaires, Physiopathologie Et Traitement Des Maladies Neurodégénératives Hereditary spastic paraplegias: clinical spectrum in Sudan, further deciphering of the molecular bases of autosomal recessive forms and new genes emerging
    [Show full text]
  • USP8 Deubiquitinates SHANK3 to Control Synapse Density and SHANK3 Activity-Dependent Protein Levels
    The Journal of Neuroscience, June 6, 2018 • 38(23):5289–5301 • 5289 Cellular/Molecular USP8 Deubiquitinates SHANK3 to Control Synapse Density and SHANK3 Activity-Dependent Protein Levels Meghan Kerrisk Campbell and XMorgan Sheng Department of Neuroscience, Genentech, South San Francisco, California 94080 Mutations or altered protein levels of SHANK3 are implicated in neurodevelopmental disorders such as Phelan–McDermid syndrome, autism spectrum disorders, and schizophrenia (Guilmatre et al., 2014). Loss of SHANK3 in mouse models results in decreased synapse density and reduction in the levels of multiple synaptic proteins (Jiang and Ehlers, 2013). The family of SHANK scaffolding molecules are among the most heavily ubiquitinated proteins at the postsynaptic density. The ubiquitin-dependent proteasome degradation of SHANK is regulated by synaptic activity and may contribute to activity-dependent synaptic remodeling (Ehlers, 2003; Shin et al., 2012).However, the identity of the specific deubiquitinating enzymes and E3 ligases that regulate SHANK ubiquitination at synapses are unknown. Here we identify USP8/UBPY as a deubiquitinating enzyme that regulates SHANK3 and SHANK1 ubiquitination and protein levels. In primary rat neurons, USP8 enhances SHANK3 and SHANK1 protein levels via deubiquitination and increases dendritic spine density. Additionally, USP8 is essential for changes in SHANK3 protein levels following synaptic activity modulation. These data identify USP8 as a key modulator of SHANK3 downstream of synaptic activity. Key words: dendritic spine; deubiquitinating enzyme; SHANK1; SHANK3; ubiquitination; USP8 Significance Statement Precise regulation of the protein levels of the postsynaptic scaffolding protein SHANK3 is essential for proper neurodevelopment. Mutations of SHANK3 have been identified in Phelan–McDermid syndrome, autism spectrum disorders, and schizophrenia (Guilmatre et al., 2014).
    [Show full text]
  • Rare Germline DICER1 Variants in Pediatric Patients with Cushing’S Disease: What Is Their Role?
    ORIGINAL RESEARCH published: 03 July 2020 doi: 10.3389/fendo.2020.00433 Rare Germline DICER1 Variants in Pediatric Patients With Cushing’s Disease: What Is Their Role? Idoia Martínez de LaPiscina 1†, Laura C. Hernández-Ramírez 2†, Nancy Portillo 1,3, Ana L. Gómez-Gila 4, Inés Urrutia 1, Rosa Martínez-Salazar 1, Alejandro García-Castaño 1, Aníbal Aguayo 1, Itxaso Rica 1,5, Sonia Gaztambide 1,6, Fabio R. Faucz 2, Margaret F. Keil 2, Maya B. Lodish 2,7, Martha Quezado 8, Nathan Pankratz 9, Prashant Chittiboina 10, Edited by: John Lane 9, Denise M. Kay 11, James L. Mills 12, Luis Castaño 1* and Madhusmita Misra, 2 1 Harvard Medical School and Constantine A. Stratakis on behalf on the Spanish Corticotroph Adenomas Collaborative Massachusetts General Hospital, 1 Section on Endocrinology, Metabolism, Nutrition and Renal Diseases, Biocruces Bizkaia Health Research Institute, Cruces United States University Hospital, UPV/EHU, CIBERER, CIBERDEM, Barakaldo, Spain, 2 Section on Endocrinology and Genetics, Eunice Reviewed by: Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Kris Ann Schultz, Bethesda, MD, United States, 3 Pediatric Endocrinology Service, Alto Deba Hospital, Arrasate, Spain, 4 Pediatric Children’s Hospitals and Clinics of Endocrinology Service, Virgen del Rocío University Hospital, Sevilla, Spain, 5 Pediatric Endocrinology Service, Cruces Minnesota, United States University Hospital, Barakaldo, Spain, 6 Endocrinology Service, Cruces University Hospital, Barakaldo,
    [Show full text]