DOCSLIB.ORG

The USP7 Protein Interaction Network and Its Roles in Tumorigenesis Ahood Al-Eidan A,B, Yihua Wang A, Paul Skipp A, Rob M

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The USP7 Protein Interaction Network and Its Roles in Tumorigenesis Ahood Al-Eidan A,B, Yihua Wang A, Paul Skipp A, Rob M + MODEL Genes & Diseases xxx (xxxx) xxx Available online at www.sciencedirect.com ScienceDirect journal homepage: http://ees.elsevier.com/gendis/default.asp REVIEW ARTICLE The USP7 protein interaction network and its roles in tumorigenesis Ahood Al-Eidan a,b, Yihua Wang a, Paul Skipp a, Rob M. Ewing a,* a School of Biological Sciences, B85 Life Sciences, University of Southampton, Southampton, SO17 1BJ, UK b Department of Biology, College of Sciences, Imam Abdulrahman Bin Faisal University, P.O. Box 1982, Dammam, Saudi Arabia Received 27 May 2020; received in revised form 14 October 2020; accepted 15 October 2020 KEYWORDS Abstract Ubiquitin-specific protease (USP7), also known as Herpesvirus-associated ubiquitin- Cancer; specific protease (HAUSP), is a deubiquitinase. There has been significant recent attention on Deubiquitinase; USP7 following the discovery that USP7 is a key regulator of the p53-MDM2 pathway. The USP7 Protein network; protein is 130 kDa in size and has multiple domains which bind to a diverse set of proteins. Proteineprotein These interactions mediate key developmental and homeostatic processes including the cell interaction; cycle, immune response, and modulation of transcription factor and epigenetic regulator ac- Proteomics; tivity and localization. USP7 also promotes carcinogenesis through aberrant activation of the USP7 Wnt signalling pathway and stabilization of HIF-1a. These findings have shown that USP7 may induce tumour progression and be a therapeutic target. Together with interest in devel- oping USP7 as a target, several studies have defined new protein interactions and the regula- tory networks within which USP7 functions. In this review, we focus on the protein interactions of USP7 that are most important for its cancer-associated roles. Copyright ª 2020, Chongqing Medical University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/ by-nc-nd/4.0/). The ubiquitin proteasome system (UPS) levels starting with the control of gene transcription and translation and finishing with the degradation of damaged, 1 Ubiquitination is a key post-translational modification gov- unnecessary or short-term proteins. The ubiquitin- erning protein turnover.1 This process is essential for the proteasome system (UPS) has been identified as a key cellular viability and homeostasis which exist at multiple regulator for the targeting of proteins especially those * Corresponding author. E-mail addresses: [email protected] (A. Al-Eidan), [email protected] (Y. Wang), [email protected] (P. Skipp), rob. [email protected] (R.M. Ewing). Peer review under responsibility of Chongqing Medical University. https://doi.org/10.1016/j.gendis.2020.10.004 2352-3042/Copyright ª 2020, Chongqing Medical University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Please cite this article as: Al-Eidan A et al., The USP7 protein interaction network and its roles in tumorigenesis, Genes & Diseases, https://doi.org/10.1016/j.gendis.2020.10.004 + MODEL 2 A. Al-Eidan et al. involved in processes such as the cell cycle, gene tran- Deubiquitinating enzymes (DUBS) scription, apoptosis and DNA repair. Through these pro- cesses, the UPS plays a role in diverse diseases such as Deubiquitinating enzymes (DUBs) oppose the E3 function- cancer, cardiovascular diseases and Alzheimer’s disease. ality by cleaving ubiquitin from from 1) ubiquitin precursor; Colorectal adenocarcinoma is a well-studied example of 2) protein substrate; 3) another ubiquitin within a poly- how important protein turnover and the UPS is in tumori- ubiquitin chain.6 There are approximately 80 DUBs which 1,2 genesis. As an example, the UPS regulates Wnt/b-cat- are classified according to the sequence homology of the enin/APC/TCF4 signalling which is implicated in the catalytic domain into six groups: ubiquitin-specific pro- proliferation of epithelial cells in the base of colonic crypts. teases (USP), ubiquitin carboxy-terminal hydrolases (UCH), The frequent mutations that alter Wnt signalling in colo- ovarian-tumour proteases (OUT), Machado-Joseph disease rectal and other solid tumours do so via stabilization of b- protein domain proteases (MJD), JAMM/MPN domain- 3 catenin leading to cell proliferation. associated metallopeptidases (JAMM) and monocyte The UPS tags proteins for destruction by the covalent chemotactic protein-induced protein (MCPIP).6 Addition- attachment of small protein ubiquitin (Ub) molecules and ally, two groups of DUBs have been recently detected; the proteasomal degradation of the tagged protein (Fig. 1). Monocyte Chemotactic Protein-Induced Proteins (MCPIPs) The ubiquitination process requires three consecutive and the MINDY family (MIU-containing novel DUB family).6 steps including the ATP-dependent activation performed Many DUBs can modify protein localization, trafficking by ubiquitin-activating (E1), ubiquitin conjugation (E2) and promote stability by deubiquitination of mono-Ub or and ubiquitin ligase (E3) enzymes of which the E3 is poly-Ub chains.6 DUBs inhibitors represent a new strategy 4 essentially in charge of substrate choice (Fig. 1). E3 of anti-cancer therapeutic and several are now in preclin- stimulates the formation of an isopeptide bond between ical stages of development7; in particular the USPs which the lysine of the target protein and the carboxyl-terminus are implicated in diverse human diseases and are the of ubiquitin. Polyubiquitination results from multiple largest group of DUBS, comprising more than 60 human ubiquitin links to a protein substrate and mono-Ub results members.6 from a single ubiquitin molecule linked to a protein sub- strate.1 In humans, there are around 600 such ubiquitin E3 ligases which are assembled into three noteworthy classes Ubiquitin-specific-peptidase 7 (USP7) characterised by the sequence homology known as the HECT, RING or RING-between-RING.1,2 Notably, aberrant The focus of this review is USP7, also known as Herpes- E3 ligases expression can function as tumour suppressors virus-Associated Ubiquitin-Specific Protease (HAUSP). We or oncogenes based on ubiquitin target substrates in focus here on the proteineprotein interactions and colorectal cancer.5 functions of USP7. Significant progress is being made on Figure 1 Key elements of the ubiquitin-proteasome system (UPS). Ubiquitin conjugation itself is mediated by the E1-E2-E3 enzymes respectively with the ubiquitin ligases (E3) targeting the ubiquitin to specific substrates. Deubiquitinase proteins (DUBs), (indicated by *) are proteases that act to cleave ubiquitin from substrate proteins and one of the most important effects of this is to antagonize the targeting of proteins for proteasomal degradation. Please cite this article as: Al-Eidan A et al., The USP7 protein interaction network and its roles in tumorigenesis, Genes & Diseases, https://doi.org/10.1016/j.gendis.2020.10.004 + MODEL USP7 protein interaction network and its functions 3 the development of therapeutics targeting USP7 and we USP7 regulation of Wnt signalling direct the reader to recent studies and reviews of this 8,9(p7) area which we will not cover here. An early study of CID-deleted APC mutants lead to accumulation of b-cat- USP7 showed that it interacts with infected cell poly- enin, activating Wnt signalling,16,17 and a role for USP7 in peptide 0 (ICP0) to activate Herpes simplex virus type 1 this has been discovered. Specifically, USP7 binds the N- 10 (HSV-1), hence the name HAUSP (in this review we refer terminus of b-catenin when the APC losses CID confirming to the protein as USP7). USP7 regulates many target the essential role of USP7 in the activation of Wnt signalling proteins and interactors through its deubiquitinating ac- and cell survival in colorectal cancers.13 Ma et al showed 11 tivity and has been shown to have an essential role in that the USP7/RNF220 complex activates the Wnt pathway the regulation of stability for proteins that are implicated while knockdown of USP7 using a small interfering RNA in crucial cellular procedures such as mitosis, apoptosis, (siRNA) strategy, reduces the expression of Wnt.18 cell cycle, DNA replication, neuronal development, and Conversely, a recent study proposed that USP7 deubiquiti- 9 epigenetic modulators. Aberrant USP7 expression was nates b-catenin and RNF220 independently and works as a 9 observed in several non-solid and solid tumours. The tumour-specific condition when APC is mutated. Addition- knockout strategy for the USP7 was lethal in mice due to ally, USP7 inhibition does not affect the Wnt activation the p53 activation which leads to a dramatic reduction in physiologically.13 12 proliferation and termination during development. USP7 Adenomatous Polyposis Coli (APC) is one of the compo- destabilizes the tumour-suppressor p53 creating interest nents of the destruction complex that regulates b-catenin 9 in it as a target in oncology and recent work suggests through phosphorylation and ubiquitination leading to its that USP7 may also be a tumour-specific drug target for degradation in the cytoplasm.19 b-catenin is the key 13 colorectal tumours with APC mutations. modulator of the Wnt signalling pathway with a mutation in APC causing an aberrant Wnt activation which is observed USP7 structure in most colorectal cancers.13 Previous studies revealed that the CID domain in the APC, which is located between the The multi-domain architecture of USP7 consists of 1102 second and third twenty amino acid repeats, is responsible 16 amino acids, and facilitates its diverse roles (Fig. 2).11,14 for inhibiting b-catenin. The CID is located in the right of USP7 contains a catalytic domain (208e560), N-terminal the mutation cluster region (MCR) which is mutated in most contains 50 amino acids followed by a TNF receptor- colorectal cancer patients. Despite the significance of the associated factor (TRAF) domain which is linked to a CID role, the DUBs role in regulating b-catenin is not fully 13 substrate peptide binding and five C-terminal ubiquitin- understood in colorectal cancer.
Load more

Recommended publications
  • USP7 [6His-Tagged] Deubiquitylating Enzyme
    USP7 [6His-tagged] Deubiquitylating Enzyme Alternate Names: Herpesvirus-Associated Ubiquitin-Specific Protease, HAUSP VMW110- associated protein Cat. No. 64-0003-050 Quantity: 50 µg Lot. No. 1736 Storage: -70˚C FOR RESEARCH USE ONLY NOT FOR USE IN HUMANS CERTIFICATE OF ANALYSIS Page 1 of 2 Background Physical Characteristics The deubiquitylating enzymes (DUBs) Species: human Protein Sequence: Please see page 2 regulate ubiquitin dependent signaling pathways. The activities of the DUBs Source: E. coli expression include the generation of free ubiquitin from precursor molecules, the recy- Quantity: 50 μg cling of ubiquitin following substrate Concentration: 0.5 mg/ml degradation to maintain cellular ubiq- uitin homeostasis and the removal Formulation: 50 mM HEPES pH 7.5, of ubiquitin or ubiquitin-like proteins 150 mM sodium chloride, 2 mM (UBL) modifications through chain dithiothreitol, 10% glycerol editing to rescue proteins from protea- somal degradation or to influence cell Molecular Weight: ~130 kDa signalling events (Komander et al., 2009). There are two main classes of Purity: >80% by InstantBlue™ SDS-PAGE DUB, cysteine proteases and metallo- Stability/Storage: 12 months at -70˚C; proteases. Ubiquitin specific process- aliquot as required ing protease 7 (USP-7) is a member of the cysteine protease enzyme fam- ily and cloning of the gene in humans Quality Assurance was first described by Everett et al. (1997). Overexpression of p53 and Purity: Protein Identification: USP7 stabilizes p53 through the re- 4-12% gradient SDS-PAGE Confirmed by mass spectrometry. InstantBlue™ staining moval of ubiquitin moieties from polyu- Lane 1: MW markers Deubiquitylating Enzyme Assay: biquitylated p53 (Kon et al., 2010).
    [Show full text]
  • Therapeutic Inhibition of USP7-PTEN Network in Chronic Lymphocytic Leukemia: a Strategy to Overcome TP53 Mutated/ Deleted Clones
    www.impactjournals.com/oncotarget/ Oncotarget, 2017, Vol. 8, (No. 22), pp: 35508-35522 Priority Research Paper Therapeutic inhibition of USP7-PTEN network in chronic lymphocytic leukemia: a strategy to overcome TP53 mutated/ deleted clones Giovanna Carrà1, Cristina Panuzzo1, Davide Torti1,2, Guido Parvis2,3, Sabrina Crivellaro1, Ubaldo Familiari4, Marco Volante4,5, Deborah Morena5, Marcello Francesco Lingua5, Mara Brancaccio6, Angelo Guerrasio1, Pier Paolo Pandolfi7, Giuseppe Saglio1,2,3, Riccardo Taulli5 and Alessandro Morotti1 1 Department of Clinical and Biological Sciences, University of Turin, San Luigi Gonzaga Hospital, Orbassano, Italy 2 Division of Internal Medicine - Hematology, San Luigi Gonzaga Hospital, Orbassano, Italy 3 Division of Hematology, Azienda Ospedaliera, Mauriziano, Torino, Italy 4 Division of Pathology, San Luigi Hospital, Orbassano, Italy 5 Department of Oncology, University of Turin, San Luigi Gonzaga Hospital, Orbassano, Italy 6 Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy 7 Cancer Genetics Program, Beth Israel Deaconess Cancer Center, Department of Medicine and Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA Correspondence to: Alessandro Morotti, email: [email protected] Correspondence to: Riccardo Taulli, email: [email protected] Keywords: chronic lymphocytic leukemia, USP7, PTEN, miR181, miR338 Received: June 14, 2016 Accepted: February 20, 2017 Published: March 17, 2017 Copyright: Carrà et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC-BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT Chronic Lymphocytic Leukemia (CLL) is a lymphoproliferative disorder with either indolent or aggressive clinical course.
    [Show full text]
  • Targeting Mdm2 and Mdmx in Cancer Therapy: Better Living Through Medicinal Chemistry?
    Subject Review Targeting Mdm2 and Mdmx in Cancer Therapy: Better Living through Medicinal Chemistry? Mark Wade and Geoffrey M.Wahl Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, California Abstract In the second model, Mdm2 and Mdmx are proposed to form Genomic and proteomic profiling of human tumor a complex that is more effective at inhibiting p53 transactiva- samples and tumor-derived cell lines are essential for tion or enhancing p53 turnover. Although the former possibility the realization of personalized therapy in oncology. has not been excluded, several studies indicate that Mdm2 Identification of the changes required for tumor initiation and Mdmx function as a heterodimeric pair to augment p53 or maintenance will likely provide new targets for degradation. Mdm2 is a member of the RING E3 ubiquitin small-molecule and biological therapeutics.For ligase family and promotes proteasome-dependent degradation example, inactivation of the p53 tumor suppressor of p53. By binding to the target substrate and to an E2 ubiquitin- pathway occurs in most human cancers.Although this conjugating enzyme, RING E3s facilitate E2-to-substrate can be due to frank p53 gene mutation, almost half of all ubiquitin transfer (5). Similar to other RING E3s, it does not cancers retain the wild-type p53 allele, indicating that seem that Mdm2 forms a covalent link with ubiquitin during the pathway is disabled by other means.Alternate the reaction. Thus, Mdm2 does not have a ‘‘classic’’ catalytic mechanisms include deletion or epigenetic inactivation site but acts as a molecular scaffold that presumably positions of the p53-positive regulator arf, methylation of the p53 p53 for E2-dependent ubiquitination (Fig.
    [Show full text]
  • USP7 Couples DNA Replication Termination to Mitotic Entry
    bioRxiv preprint doi: https://doi.org/10.1101/305318; this version posted April 20, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. USP7 couples DNA replication termination to mitotic entry Antonio Galarreta1*, Emilio Lecona1*, Pablo Valledor1, Patricia Ubieto1,2, Vanesa Lafarga1, Julia Specks1 & Oscar Fernandez-Capetillo1,3 1Genomic Instability Group, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain 2Current Address: DNA Replication Group, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain 3Science for Life Laboratory, Division of Genome Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, S-171 21 Stockholm, Sweden *Co-first authors Correspondence: E.L. ([email protected]) or O.F. ([email protected]) Lead Contact: Oscar Fernandez-Capetillo Spanish National Cancer Research Centre (CNIO) Melchor Fernandez Almagro, 3 Madrid 28029, Spain Tel.: +34.91.732.8000 Ext: 3480 Fax: +34.91.732.8028 Email: [email protected] KEYWORDS: USP7; CDK1; DNA REPLICATION; MITOSIS; S/M TRANSITION. bioRxiv preprint doi: https://doi.org/10.1101/305318; this version posted April 20, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. USP7 coordinates the S/M transition 2 SUMMARY To ensure a faithful segregation of chromosomes, DNA must be fully replicated before mitotic entry. However, how cells sense the completion of DNA replication and to what extent this is linked to the activation of the mitotic machinery remains poorly understood. We previously showed that USP7 is a replisome-associated deubiquitinase with an essential role in DNA replication.
    [Show full text]
  • The Role of Ubiquitination in NF-Κb Signaling During Virus Infection
    viruses Review The Role of Ubiquitination in NF-κB Signaling during Virus Infection Kun Song and Shitao Li * Department of Microbiology and Immunology, Tulane University, New Orleans, LA 70112, USA; [email protected] * Correspondence: [email protected] Abstract: The nuclear factor κB (NF-κB) family are the master transcription factors that control cell proliferation, apoptosis, the expression of interferons and proinflammatory factors, and viral infection. During viral infection, host innate immune system senses viral products, such as viral nucleic acids, to activate innate defense pathways, including the NF-κB signaling axis, thereby inhibiting viral infection. In these NF-κB signaling pathways, diverse types of ubiquitination have been shown to participate in different steps of the signal cascades. Recent advances find that viruses also modulate the ubiquitination in NF-κB signaling pathways to activate viral gene expression or inhibit host NF-κB activation and inflammation, thereby facilitating viral infection. Understanding the role of ubiquitination in NF-κB signaling during viral infection will advance our knowledge of regulatory mechanisms of NF-κB signaling and pave the avenue for potential antiviral therapeutics. Thus, here we systematically review the ubiquitination in NF-κB signaling, delineate how viruses modulate the NF-κB signaling via ubiquitination and discuss the potential future directions. Keywords: NF-κB; polyubiquitination; linear ubiquitination; inflammation; host defense; viral infection Citation: Song, K.; Li, S. The Role of 1. Introduction Ubiquitination in NF-κB Signaling The nuclear factor κB (NF-κB) is a small family of five transcription factors, including during Virus Infection. Viruses 2021, RelA (also known as p65), RelB, c-Rel, p50 and p52 [1].
    [Show full text]
  • Tumor Suppressors in Chronic Lymphocytic Leukemia: from Lost Partners to Active Targets
    cancers Review Tumor Suppressors in Chronic Lymphocytic Leukemia: From Lost Partners to Active Targets 1, 1, 2 1 Giacomo Andreani y , Giovanna Carrà y , Marcello Francesco Lingua , Beatrice Maffeo , 3 2, 1, , Mara Brancaccio , Riccardo Taulli y and Alessandro Morotti * y 1 Department of Clinical and Biological Sciences, University of Torino, 10043 Orbassano, Italy; [email protected] (G.A.); [email protected] (G.C.); beatrice.maff[email protected] (B.M.) 2 Department of Oncology, University of Torino, 10043 Orbassano, Italy; [email protected] (M.F.L.); [email protected] (R.T.) 3 Department of Molecular Biotechnology and Health Sciences, University of Torino, 10126 Turin, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-011-9026305 These authors equally contributed to the work. y Received: 21 January 2020; Accepted: 4 March 2020; Published: 9 March 2020 Abstract: Tumor suppressors play an important role in cancer pathogenesis and in the modulation of resistance to treatments. Loss of function of the proteins encoded by tumor suppressors, through genomic inactivation of the gene, disable all the controls that balance growth, survival, and apoptosis, promoting cancer transformation. Parallel to genetic impairments, tumor suppressor products may also be functionally inactivated in the absence of mutations/deletions upon post-transcriptional and post-translational modifications. Because restoring tumor suppressor functions remains the most effective and selective approach to induce apoptosis in cancer, the dissection of mechanisms of tumor suppressor inactivation is advisable in order to further augment targeted strategies. This review will summarize the role of tumor suppressors in chronic lymphocytic leukemia and attempt to describe how tumor suppressors can represent new hopes in our arsenal against chronic lymphocytic leukemia (CLL).
    [Show full text]
  • Análise Integrativa De Perfis Transcricionais De Pacientes Com
    UNIVERSIDADE DE SÃO PAULO FACULDADE DE MEDICINA DE RIBEIRÃO PRETO PROGRAMA DE PÓS-GRADUAÇÃO EM GENÉTICA ADRIANE FEIJÓ EVANGELISTA Análise integrativa de perfis transcricionais de pacientes com diabetes mellitus tipo 1, tipo 2 e gestacional, comparando-os com manifestações demográficas, clínicas, laboratoriais, fisiopatológicas e terapêuticas Ribeirão Preto – 2012 ADRIANE FEIJÓ EVANGELISTA Análise integrativa de perfis transcricionais de pacientes com diabetes mellitus tipo 1, tipo 2 e gestacional, comparando-os com manifestações demográficas, clínicas, laboratoriais, fisiopatológicas e terapêuticas Tese apresentada à Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo para obtenção do título de Doutor em Ciências. Área de Concentração: Genética Orientador: Prof. Dr. Eduardo Antonio Donadi Co-orientador: Prof. Dr. Geraldo A. S. Passos Ribeirão Preto – 2012 AUTORIZO A REPRODUÇÃO E DIVULGAÇÃO TOTAL OU PARCIAL DESTE TRABALHO, POR QUALQUER MEIO CONVENCIONAL OU ELETRÔNICO, PARA FINS DE ESTUDO E PESQUISA, DESDE QUE CITADA A FONTE. FICHA CATALOGRÁFICA Evangelista, Adriane Feijó Análise integrativa de perfis transcricionais de pacientes com diabetes mellitus tipo 1, tipo 2 e gestacional, comparando-os com manifestações demográficas, clínicas, laboratoriais, fisiopatológicas e terapêuticas. Ribeirão Preto, 2012 192p. Tese de Doutorado apresentada à Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo. Área de Concentração: Genética. Orientador: Donadi, Eduardo Antonio Co-orientador: Passos, Geraldo A. 1. Expressão gênica – microarrays 2. Análise bioinformática por module maps 3. Diabetes mellitus tipo 1 4. Diabetes mellitus tipo 2 5. Diabetes mellitus gestacional FOLHA DE APROVAÇÃO ADRIANE FEIJÓ EVANGELISTA Análise integrativa de perfis transcricionais de pacientes com diabetes mellitus tipo 1, tipo 2 e gestacional, comparando-os com manifestações demográficas, clínicas, laboratoriais, fisiopatológicas e terapêuticas.
    [Show full text]
  • Structural Insight Into the Mechanisms of Activation and Substrate
    University of Connecticut OpenCommons@UConn Doctoral Dissertations University of Connecticut Graduate School 12-12-2016 Structural Insight into the Mechanisms of Activation and Substrate Specificity of Human Deubiquitinating Enzyme USP7 Alexandra Pozhidaeva University of Connecticut, [email protected] Follow this and additional works at: https://opencommons.uconn.edu/dissertations Recommended Citation Pozhidaeva, Alexandra, "Structural Insight into the Mechanisms of Activation and Substrate Specificity of Human Deubiquitinating Enzyme USP7" (2016). Doctoral Dissertations. 1287. https://opencommons.uconn.edu/dissertations/1287 Structural Insight into the Mechanisms of Activation and Substrate Specificity of Human Deubiquitinating Enzyme USP7 Alexandra Pozhidaeva, PhD University of Connecticut, 2016 The major part of this thesis describes studies of human ubiquitin-specific protease 7 (USP7), a deubiquitinating enzyme that regulates cellular levels of key oncoproteins and tumor suppressors. Inactivation of USP7 has recently emerged as a new approach to treatment of malignancies. However, design of potent and specific small-molecule compounds requires detailed understanding of the molecular mechanisms of USP7 substrate recognition and regulation of its catalytic activity. The goal of this work was to explore these mechanisms using solution nuclear magnetic resonance spectroscopy in combination with other methods. In our studies of USP7 substrate recognition, we structurally characterized its interaction with ICP0 protein from Herpes Simplex virus 1 and identified a novel USP7 substrate-binding site harbored within its C-terminal region. To address the question of USP7 activity regulation, we investigated its interaction with ubiquitin, which was believed to cause structural rearrangement of USP7 active site from an unproductive to a catalytically competent conformation. Surprisingly, we showed that in solution USP7 – ubiquitin interaction alone is not sufficient for activation of the enzyme as was previously postulated.
    [Show full text]
  • MYRF Is a Membrane-Associated Transcription Factor That Autoproteolytically Cleaves to Directly Activate Myelin Genes
    MYRF Is a Membrane-Associated Transcription Factor That Autoproteolytically Cleaves to Directly Activate Myelin Genes Helena Bujalka1., Matthias Koenning1., Stacey Jackson1, Victoria M. Perreau1, Bernard Pope2, Curtis M. Hay1, Stanlislaw Mitew1, Andrew F. Hill3, Q. Richard Lu4, Michael Wegner5, Rajini Srinivasan6, John Svaren6, Melanie Willingham1, Ben A. Barres7, Ben Emery1,8* 1 Department of Anatomy and Neuroscience and the Centre for Neuroscience Research, University of Melbourne, Parkville, Australia, 2 Department of Computing and Information Systems and Victorian Life Sciences Computation Initiative (VLSCI), University of Melbourne, Parkville, Australia, 3 Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Australia, 4 Department of Developmental Biology and Kent Waldrep Foundation Center for Basic Neuroscience Research on Nerve Growth and Regeneration, University of Texas at Austin, Austin, Texas, United States of America, 5 Institute for Biochemistry, University Erlangen-Nuernberg, Germany, 6 Waisman Center, University of Wisconsin–Madison, Madison, Wisconsin, United States of America, 7 Department of Neurobiology, Stanford University, Stanford, California, United States of America, 8 Florey Institute of Neuroscience and Mental Health, Parkville, Australia Abstract The myelination of axons is a crucial step during vertebrate central nervous system (CNS) development, allowing for rapid and energy efficient saltatory conduction of nerve impulses. Accordingly, the differentiation of oligodendrocytes, the myelinating cells of the CNS, and their expression of myelin genes are under tight transcriptional control. We previously identified a putative transcription factor, Myelin Regulatory Factor (Myrf), as being vital for CNS myelination. Myrf is required for the generation of CNS myelination during development and also for its maintenance in the adult.
    [Show full text]
  • Sexual Dimorphism in Brain Transcriptomes of Amami Spiny Rats (Tokudaia Osimensis): a Rodent Species Where Males Lack the Y Chromosome Madison T
    Ortega et al. BMC Genomics (2019) 20:87 https://doi.org/10.1186/s12864-019-5426-6 RESEARCHARTICLE Open Access Sexual dimorphism in brain transcriptomes of Amami spiny rats (Tokudaia osimensis): a rodent species where males lack the Y chromosome Madison T. Ortega1,2, Nathan J. Bivens3, Takamichi Jogahara4, Asato Kuroiwa5, Scott A. Givan1,6,7,8 and Cheryl S. Rosenfeld1,2,8,9* Abstract Background: Brain sexual differentiation is sculpted by precise coordination of steroid hormones during development. Programming of several brain regions in males depends upon aromatase conversion of testosterone to estrogen. However, it is not clear the direct contribution that Y chromosome associated genes, especially sex- determining region Y (Sry), might exert on brain sexual differentiation in therian mammals. Two species of spiny rats: Amami spiny rat (Tokudaia osimensis) and Tokunoshima spiny rat (T. tokunoshimensis) lack a Y chromosome/Sry, and these individuals possess an XO chromosome system in both sexes. Both Tokudaia species are highly endangered. To assess the neural transcriptome profile in male and female Amami spiny rats, RNA was isolated from brain samples of adult male and female spiny rats that had died accidentally and used for RNAseq analyses. Results: RNAseq analyses confirmed that several genes and individual transcripts were differentially expressed between males and females. In males, seminal vesicle secretory protein 5 (Svs5) and cytochrome P450 1B1 (Cyp1b1) genes were significantly elevated compared to females, whereas serine (or cysteine) peptidase inhibitor, clade A, member 3 N (Serpina3n) was upregulated in females. Many individual transcripts elevated in males included those encoding for zinc finger proteins, e.g.
    [Show full text]
  • 1 Mutational Heterogeneity in Cancer Akash Kumar a Dissertation
    Mutational Heterogeneity in Cancer Akash Kumar A dissertation Submitted in partial fulfillment of requirements for the degree of Doctor of Philosophy University of Washington 2014 June 5 Reading Committee: Jay Shendure Pete Nelson Mary Claire King Program Authorized to Offer Degree: Genome Sciences 1 University of Washington ABSTRACT Mutational Heterogeneity in Cancer Akash Kumar Chair of the Supervisory Committee: Associate Professor Jay Shendure Department of Genome Sciences Somatic mutation plays a key role in the formation and progression of cancer. Differences in mutation patterns likely explain much of the heterogeneity seen in prognosis and treatment response among patients. Recent advances in massively parallel sequencing have greatly expanded our capability to investigate somatic mutation. Genomic profiling of tumor biopsies could guide the administration of targeted therapeutics on the basis of the tumor’s collection of mutations. Central to the success of this approach is the general applicability of targeted therapies to a patient’s entire tumor burden. This requires a better understanding of the genomic heterogeneity present both within individual tumors (intratumoral) and amongst tumors from the same patient (intrapatient). My dissertation is broadly organized around investigating mutational heterogeneity in cancer. Three projects are discussed in detail: analysis of (1) interpatient and (2) intrapatient heterogeneity in men with disseminated prostate cancer, and (3) investigation of regional intratumoral heterogeneity in
    [Show full text]
  • An Allosteric Interaction Links USP7 to Deubiquitination and Chromatin Targeting of UHRF1 Zhi-Min Zhang University of California, Riverside
    University of the Pacific Scholarly Commons College of the Pacific aF culty Articles All Faculty Scholarship 9-1-2015 An Allosteric Interaction Links USP7 to Deubiquitination and Chromatin Targeting of UHRF1 Zhi-Min Zhang University of California, Riverside Scott .B Rothbart Van Andel Research Institute David F. Allison University of North Carolina at Chapel Hill Qian Cai University of California, Riverside Joseph S. Harrison University of the Pacific, [email protected] See next page for additional authors Follow this and additional works at: https://scholarlycommons.pacific.edu/cop-facarticles Part of the Biochemistry, Biophysics, and Structural Biology Commons, and the Chemistry Commons Recommended Citation Zhang, Z., Rothbart, S. B., Allison, D. F., Cai, Q., Harrison, J. S., Li, L., Wang, Y., Strahl, B. D., Wang, G. G., & Song, J. (2015). An Allosteric Interaction Links USP7 to Deubiquitination and Chromatin Targeting of UHRF1. Cell Reports, 12(9), 1400–1406. DOI: 10.1016/j.celrep.2015.07.046 https://scholarlycommons.pacific.edu/cop-facarticles/575 This Article is brought to you for free and open access by the All Faculty Scholarship at Scholarly Commons. It has been accepted for inclusion in College of the Pacific aF culty Articles by an authorized administrator of Scholarly Commons. For more information, please contact [email protected]. Authors Zhi-Min Zhang, Scott .B Rothbart, David F. Allison, Qian Cai, Joseph S. Harrison, Lin Li, Yinsheng Wang, Brian D. Strahl, Gang Greg Wang, and Jikui Song This article is available at Scholarly Commons: https://scholarlycommons.pacific.edu/cop-facarticles/575 Report An Allosteric Interaction Links USP7 to Deubiquitination and Chromatin Targeting of UHRF1 Graphical Abstract Authors Zhi-Min Zhang, Scott B.
    [Show full text]