DOCSLIB.ORG

Correlation of NEDD4-1 and PTEN Expression with the Invasive Capacity of Pituitary Adenomas

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Correlation of NEDD4-1 and PTEN Expression with the Invasive Capacity of Pituitary Adenomas 96 MOLECULAR AND CLINICAL ONCOLOGY 6: 96-100, 2017 Correlation of NEDD4-1 and PTEN expression with the invasive capacity of pituitary adenomas JING ZHANG1, XIN LI2 and YANYAN ZHANG1 1Department of Neurosurgery, Heze Municipal Hospital, Heze, Shandong 274000; 2Department of Neurosurgery, Liaocheng �eople's Hospital and Liaocheng Clinical School of Taishan Medical University, Liaocheng, Shandong 252000, �.R. China Received December 21, 2015; Accepted November 3, 2016 DOI: 10.3892/mco.2016.1092 Abstract. The aim of the present study was to correlate the have advanced the current knowledge on the pathogenesis expression of neural precursor cell expressed developmentally of pituitary adenoma, the precise mechanisms and causes downregulated 4-1 (NEDD4-1) and phosphatase and tensin have remained elusive. �hosphatase and tensin homolog homolog deleted on chromosome 10 (���������������������TEN) with the inva- deleted on chromosome 10 (PTEN), the first known tumor sive capacity of pituitary adenomas. A total of 50 pituitary suppressor gene with phosphatase activity, was first cloned adenoma tissues and 10 normal pituitary tissues were divided and named in 1997 (2,3). In 2007, neural precursor cell into the invasive group (26 cases), the non-invasive group expressed developmentally down regulated 4-1 (NEDD4-1) (24 cases) and the normal group (10 cases). The expression of was identified by Wang et al (4). NEDD4-1 has an important NEDD4-1 and �TEN was determined by immunohistochem- role in downregulating the levels of �TEN. It can promote istry. NEDD4-1 was revealed to be located in the nucleus and the degradation of �TEN in the proteasome by adding a cytoplasm, whereas �TEN was only located in the cytoplasm. molecular marker, ubiquitin (4-8). To the best of our knowl- Furthermore, expression of NEDD4-1 was higher in pituitary edge, no previous studies have assessed whether NEDD4-1 adenomas compared with normal pituitary tissues (�<0.05), is expressed in pituitary adenomas or whether its expression and higher in the invasive group compared with the non-inva- is associated with their invasiveness or with �TEN. In the sive group, whereas the opposite trend was observed for �TEN. present study, the expression of NEDD4-1 and �TEN in There was a strong negative correlation between NEDD4-1 and invasive and non-invasive pituitary adenomas and normal �TEN expression, indicating a dependency between the two pituitary tissues were analyzed using immunohistochem- and an association with invasiveness. In conclusion, NEDD4-1 istry, and their correlation was assessed. The present study may serve as a diagnostic and prognostic factor, and as a novel aimed to investigate the possible molecular mechanisms of therapeutic target, in pituitary adenomas. the genesis and invasiveness of pituitary adenomas, and to identify biomarkers for early detection as well as character- Introduction ization of the biological behavior of pituitary adenomas. �ituitary adenoma is a common type of neurological Materials and methods neoplasm and the third most common type of intracranial tumors. While pituitary adenomas are classified as benign Patients and reagents. The specimens for immunohistochem- tumors, a proportion of them exhibit invasive growth towards ical analysis were collected from 50 patients with pituitary the surrounding normal structures, including the skull, adenomas who underwent treatment at the Department of cavernous sinus, dura, sphenoid bone, the third ventricle, and Neurosurgery of Heze Municipal Hospital (Heze, China) even the pharyngonasal cavity (1). Although recent studies from July 2012 to May 2015. The patient cohort comprised 27 males and 23 females, with a male-to-female ratio of 1.17:1. The average age of the patients was 38.7 years (range, 18-78 years). According to the presence or absence of inva- sion of the tumors to the surrounding bones and cavernous Correspondence to: Dr Jing Zhang, Department of Neurosurgery, sinus, patients were divided into the invasive group Heze Municipal Hospital, 2888 Caozhou Road, Heze, Shandong 274000, �.R. China (26 cases) and the non‑invasive group (24 cases) (Table Ⅰ). E-mail: [email protected] Furthermore, 10 samples of normal pituitary tissues were obtained through normal autopsy. Rabbit anti-human Key words: invasion, neural precursor cells, developmentally NEDD4-1 polyclonal antibody (cat. no. bs-3506R-AF350; downregulated 4-1, phosphatase and tensin homolog deleted on 1:200 dilution) and rabbit anti-human �TEN polyclonal anti- chromosome 10, pituitary adenomas, immunohistochemistry body (cat. no. DM-0748R,1:300 dilution) were from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), and secondary antibodies were from a designated immunohistochemistry ZHANG et al: EX�RESSION AND CORRELATION OF NEDD4-1 AND �TEN IN �ITUITARY ADENOMAS 97 kit for rabbit antibodies (Beijing Zhongshan Golden Bridge Table I. Clinical data of patients with pituitary adenoma. Biotechnology Co., Ltd., China). All experiments and use of the specimens in the present study were approved by Invasive Non-invasive the Ethics Committee of Heze Municipal Hospital (Heze, Characteristic group (n) group (n) China), and all patients provided written informed consent prior to enrollment. Cases 26 24 Age (years) Immunohistochemical staining procedures. ≥40 20 15 �araffin-embedded specimens were cut into 4-µm thick <40 6 9 sections, which were deparaffinized in turpentine, rehy- Gender drated in an ascending ethanol series and incubated in citrate Male 13 14 buffer (0.01 mol/l, pH 6.0) with water bath heating (~98˚C) Female 13 10 for 15 min for antigen recovery. Following a general strep- tavidin-biotin complex immunohistochemical protocol (9), Endocrine type immunoreactivity was visualized with diaminobenzidine and ACTH 2 5 the nuclei were counterstained with hematoxylin. Sections GH 6 3 were then subsequently dehydrated in a series of graded alco- �RL 8 10 hols, cleared in xylene and mounted. �rostate cancer samples Others 10 6 were used as the positive control and slides incubated with Operative method phosphate-buffered saline without antibody were used as the Transcranial 21 6 negative control. Trans-sphenoidal 5 18 Evaluation of immunohistochemical staining. For each ACTH, adrenocorticotropic hormone; GH, growth hormone; �RL, sample, 10 microscopic fields (magnification, x200) were prolactin. selected from the tissue sections with the strongest immune response. A total of 100 cells per field were counted and averaged. The percentage of positively stained cells was recorded. Staining was rated according to the color intensity and the percentage of positive cells. �ositive staining was and/or the nucleus (Fig. 1). The positive rate of NEDD4-1 defined as the staining intensity being markedly higher than expression in the normal control group [10.0% (1/10)] was that of the background. Staining was classified into four significantly lower compared with that in the experimental categories according to the number of stained cells and the group [52.0% (26/50); P<0.05] (Table II). The positive rate staining intensity: Negative [- (i.e., the staining intensity was of NEDD4-1 was 37.5% (9/24) in the non-invasive pituitary not significantly different from the background)]; weakly adenomas group and 65.3% (17/26) in the invasive adenomas positive [+ (i.e., the percentage of positively stained cells group, thereby revealing a significantly increasing trend with was <10%, or the staining intensity was weakly positive or the increasing degree of tumor invasiveness (�<0.05); the only individual cells were positive to strongly positive)]; differences were also significant compared with the control moderately positive [++ (i.e., the percentage of positive cells group (�<0.05). was 10‑60%)]; strongly positive [+++ (i.e., the percentage of positively stained cells was >60%, or the staining intensity PTEN protein expression in pituitary tissues/adenoma was moderately to strongly positive and only a few cells were decreases with increasing degree of malignancy. Immuno- weakly positive]. The known positive prostate section was histochemistry revealed that �TEN was mainly expressed in selected as the positive control. the cytoplasm, displaying as a brownish-yellow stain (Fig. 2). The rate of �TEN expression in the normal control group Statistical analysis. S�SS 18.0 software was used for statis- [100.0% (10/10)] was significantly higher compared with that in tical analysis. The two-samples t-test and analysis of variance the experimental group [54.0% (27/50); P<0.05] (Table III). were used to assess differences between groups. The χ2 test The positive rate of �TEN was 83.3% (20/24) in the non-inva- and exact test on a four-fold table were used to determine sive pituitary adenoma group and 26.9% (7/26) in the invasive differences in the positive staining rate between normal pitu- pituitary adenoma group, thereby revealing a significantly itary tissues and invasive, as well as non-invasive, pituitary decreasing trend with the increasing degree of tumor invasive- adenomas. The Line x List χ2 test and the one-sided test were ness (P<0.05); the differences were also significant compared used for correlation analyses. α=0.05 was considered as the with the control group (�<0.05). level of significance. P<0.05 was considered to indicate a statistically significant difference. Inverse correlation of NEDD4‑1 and PTEN protein expres‑ sion in pituitary adenomas. Among the 26 cases of pituitary Results adenomas which stained positive for NEDD4-1, four were posi- tive for �TEN. Furthermore, among the 27 cases of pituitary NEDD4‑1 expression in pituitary tissues/adenoma increases adenomas which stained positive for �TEN, 24 were negative with the degree of malignancy. Immunohistochemistry for NEDD4-1. Statistical analysis revealed a negative correla- revealed that NEDD4-1 was expressed in the cytoplasm tion between the two proteins (γ=-0.711, �<0.05) (Table IV). 98 MOLECULAR AND CLINICAL ONCOLOGY 6: 96-100, 2017 A B C Figure 1. NEDD4‑1 expression in normal pituitary tissue and pituitary adenomas (magnification, x200). NEDD4‑1 was expressed in the cytoplasm and/or the nucleus.
Load more

Recommended publications
  • The Role of the Ubiquitin Ligase Nedd4-1 in Skeletal Muscle Atrophy
    The Role of the Ubiquitin Ligase Nedd4-1 in Skeletal Muscle Atrophy by Preena Nagpal A thesis submitted in conformity with the requirements for the degree of Masters in Medical Science Institute of Medical Science University of Toronto © Copyright by Preena Nagpal 2012 The Role of the Ubiquitin Ligase Nedd4-1 in Skeletal Muscle Atrophy Preena Nagpal Masters in Medical Science Institute of Medical Science University of Toronto 2012 Abstract Skeletal muscle (SM) atrophy complicates many illnesses, diminishing quality of life and increasing disease morbidity, health resource utilization and health care costs. In animal models of muscle atrophy, loss of SM mass results predominantly from ubiquitin-mediated proteolysis and ubiquitin ligases are the key enzymes that catalyze protein ubiquitination. We have previously shown that ubiquitin ligase Nedd4-1 is up-regulated in a rodent model of denervation- induced SM atrophy and the constitutive expression of Nedd4-1 is sufficient to induce myotube atrophy in vitro, suggesting an important role for Nedd4-1 in the regulation of muscle mass. In this study we generate a Nedd4-1 SM specific-knockout mouse and demonstrate that the loss of Nedd4-1 partially protects SM from denervation-induced atrophy confirming a regulatory role for Nedd4-1 in the maintenance of muscle mass in vivo. Nedd4-1 did not signal downstream through its known substrates Notch-1, MTMR4 or FGFR1, suggesting a novel substrate mediates Nedd4-1’s induction of SM atrophy. ii Acknowledgments and Contributions I would like to thank my supervisor, Dr. Jane Batt, for her undying support throughout my time in the laboratory.
    [Show full text]
  • Nedd4 and Nedd4-2: Closely Related Ubiquitin-Protein Ligases with Distinct Physiological Functions
    Cell Death and Differentiation (2010) 17, 68–77 & 2010 Macmillan Publishers Limited All rights reserved 1350-9047/10 $32.00 www.nature.com/cdd Review Nedd4 and Nedd4-2: closely related ubiquitin-protein ligases with distinct physiological functions B Yang*,1 and S Kumar*,2 The Nedd4 (neural precursor cell-expressed developmentally downregulated gene 4) family of ubiquitin ligases (E3s) is characterized by a distinct modular domain architecture, with each member consisting of a C2 domain, 2–4 WW domains, and a HECT-type ligase domain. Of the nine mammalian members of this family, Nedd4 and its close relative, Nedd4-2, represent the ancestral ligases with strong similarity to the yeast, Rsp5. In Saccharomyces cerevisiae Rsp5 has a key role in regulating the trafficking, sorting, and degradation of a large number of proteins in multiple cellular compartments. However, in mammals the Nedd4 family members, including Nedd4 and Nedd4-2, appear to have distinct functions, thereby suggesting that these E3s target specific proteins for ubiquitylation. In this article we focus on the biology and emerging functions of Nedd4 and Nedd4-2, and review recent in vivo studies on these E3s. Cell Death and Differentiation (2010) 17, 68–77; doi:10.1038/cdd.2009.84; published online 26 June 2009 Ubiquitylation controls biological signaling in many different ubiquitin-protein ligase (E3). A protein can be monoubiquity- ways.1,2 For example, the ubiquitylation of a misfolded or lated, multi-monoubiquitylated, or polyubiquitylated, and the damaged protein leads to its degradation by the 26S type of ubiquitylation determines the fate of the protein.1,2 proteasome before it can get to its subcellular site where it Ubiquitin itself contains seven lysine residues, all of which can normally functions.
    [Show full text]
  • E3 Ubiquitin Ligases: Key Regulators of Tgfβ Signaling in Cancer Progression
    International Journal of Molecular Sciences Review E3 Ubiquitin Ligases: Key Regulators of TGFβ Signaling in Cancer Progression Abhishek Sinha , Prasanna Vasudevan Iyengar and Peter ten Dijke * Department of Cell and Chemical Biology and Oncode Institute, Leiden University Medical Center, 2300 RC Leiden, The Netherlands; [email protected] (A.S.); [email protected] (P.V.I.) * Correspondence: [email protected]; Tel.: +31-71-526-9271 Abstract: Transforming growth factor β (TGFβ) is a secreted growth and differentiation factor that influences vital cellular processes like proliferation, adhesion, motility, and apoptosis. Regulation of the TGFβ signaling pathway is of key importance to maintain tissue homeostasis. Perturbation of this signaling pathway has been implicated in a plethora of diseases, including cancer. The effect of TGFβ is dependent on cellular context, and TGFβ can perform both anti- and pro-oncogenic roles. TGFβ acts by binding to specific cell surface TGFβ type I and type II transmembrane receptors that are endowed with serine/threonine kinase activity. Upon ligand-induced receptor phosphorylation, SMAD proteins and other intracellular effectors become activated and mediate biological responses. The levels, localization, and function of TGFβ signaling mediators, regulators, and effectors are highly dynamic and regulated by a myriad of post-translational modifications. One such crucial modification is ubiquitination. The ubiquitin modification is also a mechanism by which crosstalk with other signaling pathways is achieved. Crucial effector components of the ubiquitination cascade include the very diverse family of E3 ubiquitin ligases. This review summarizes the diverse roles of E3 ligases that act on TGFβ receptor and intracellular signaling components.
    [Show full text]
  • Isolation and Characterization of the Pin1/Ess1p Homologue in Schizosaccharomyces Pombe
    RESEARCH ARTICLE 3779 Isolation and characterization of the Pin1/Ess1p homologue in Schizosaccharomyces pombe Han-kuei Huang1, Susan L. Forsburg1, Ulrik P. John2, Matthew J. O’Connell2,3 and Tony Hunter1,* 1Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA 2Trescowthick Research Laboratories, Peter MacCallum Cancer Institute, Locked Bag 1, A’Beckett Street, Melbourne, VIC 8006, Australia 3Department of Genetics University of Melbourne, Parkville, VIC 3052, Australia *Author for correspondence (e-mail: [email protected]) Accepted 13 July 2001 Journal of Cell Science 114, 3779-3788 (2001) © The Company of Biologists Ltd SUMMARY Pin1/Ess1p is a highly conserved WW domain-containing to the cyclophilin inhibitor, cyclosporin A, suggesting that peptidyl-prolyl isomerase (PPIase); its WW domain binds cyclophilin family PPIases have overlapping functions with specifically to phospho-Ser/Thr-Pro sequences and its the Pin1p PPIase. Deletion of pin1+ did not affect the DNA catalytic domain isomerizes phospho-Ser/Thr-Pro bonds. replication checkpoint, but conferred a modest increase in Pin1 PPIase activity can alter protein conformation in a UV sensitivity. Furthermore, the pin1∆ allele caused a phosphorylation-dependent manner and/or promote synthetic growth defect when combined with either cdc25- protein dephosphorylation. Human Pin1 interacts with 22 or wee1-50 but not the cdc24-1 temperature-sensitive mitotic phosphoproteins, such as NIMA, Cdc25 and Wee1, mutant. The pin1∆ strain showed increased sensitivity to and inhibits G2/M progression in Xenopus extracts. the PP1/PP2A family phosphatase inhibitor, okadaic Depletion of Pin1 in HeLa cells and deletion of ESS1 in S. acid, suggesting that Pin1p plays a role in protein cerevisiae result in mitotic arrest.
    [Show full text]
  • NEDD4 Ubiquitinates TRAF3 to Promote CD40-Mediated AKT Activation
    ARTICLE Received 5 Dec 2013 | Accepted 25 Jun 2014 | Published 29 Jul 2014 DOI: 10.1038/ncomms5513 NEDD4 ubiquitinates TRAF3 to promote CD40-mediated AKT activation Di-Feng Fang1,*, Kun He1,*, Na Wang1,*, Zhi-Hong Sang1, Xin Qiu1, Guang Xu1, Zhao Jian1, Bing Liang1, Tao Li 1, Hui-Yan Li1, Ai-Ling Li1, Tao Zhou1, Wei-Li Gong1, Baoli Yang2, Michael Karin3, Xue-Min Zhang1 & Wei-Hua Li1 CD40, a member of tumour necrosis factor receptor (TNFR) superfamily, has a pivotal role in B-cell-mediated immunity through various effector pathways including AKT kinase, but the signal transduction of CD40-meidated AKT activation is poorly understood. Here we report that the neural precursor cell expressed developmentally downregulated protein 4 (NEDD4), homologous to E6-AP Carboxyl Terminus family E3 ubiquitin ligase, is a novel component of the CD40 signalling complex. It has a key role in CD40-mediated AKT activation and is involved in modulating immunoglobulin class switch through regulating the expression of activation-induced cytidine deaminase. NEDD4 constitutively interacts with CD40 and mediates K63-linked ubiquitination of TNFR-associated factor3 (TRAF3). The ubiquitination of TRAF3 by NEDD4 is critical for CD40-mediated AKT activation. Thus, NEDD4 is a previously unknown component of the CD40 signalling complex necessary for AKT activation. 1 State Key Laboratory of Proteomics, National Center of Biomedical Analysis, Institute of Basic Medical Sciences, Beijing 100850, China. 2 Department of Obstetrics and Gynecology, Carver College of Medicine, University of Iowa, Iowa City, Iowa 52242, USA. 3 Laboratory of Gene Regulation and Signal Transduction, Cancer Center, Departments of Pharmacology and Pathology, University of California, San Diego, California 93093, USA.
    [Show full text]
  • Functional Interaction Between Pten and Nedd4 In
    FUNCTIONAL INTERACTION BETWEEN PTEN AND NEDD4 IN IGF SIGNALING A Dissertation Presented to the Faculty of Weill Cornell Graduate School of Medical Sciences in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy By Yuji Shi January 2015 © 2014 Yuji Shi FUNCTIONAL INTERACTION BETWEEN PTEN AND NEDD4 IN IGF SIGNALING Yuji Shi, Ph.D. Cornell University 2015 PTEN is a master regulator of multiple cellular processes and a potent tumor suppressor. Its biological function is mainly attributed to its lipid phosphatase activity that negatively regulates the PI3K-AKT signaling pathway. A fundamental and highly debated question remains whether PTEN can also function as a protein phosphatase in cells. This study demonstrates that PTEN is a protein tyrosine phosphatase that selectively dephosphorylates insulin receptor substrate-1 (IRS1), a mediator for transduction of insulin and IGF1 signaling. IGF signaling is defective in cells lacking NEDD4, a PTEN ubiquitin ligase, whereas AKT activation triggered by EGF or serum is unimpaired in these cells. Surprisingly, the defect of IGF signaling caused by NEDD4 deletion, including the of phosphorylation of IRS1, upstream of PI3K, can be rescued by PTEN ablation, suggesting PTEN may be a protein phosphatase for IRS1. The nature of PTEN as an IRS1 phosphatase is demonstrated by direct biochemical analysis and confirmed by cellular reconstitution. Further, we find that NEDD4 supports insulin-mediated glucose metabolism, and is required for the proliferation of IGF1 receptor (IGF1R)-dependent but not EGFR-dependent tumor cells. Taken together, PTEN is a protein phosphatase for IRS1, and its antagonism by the ubiquitin ligase NEDD4 promotes IGF/insulin signaling.
    [Show full text]
  • Herpes Simplex Virus-1 Pul56 Degrades GOPC to Alter the Plasma
    bioRxiv preprint doi: https://doi.org/10.1101/729343; this version posted August 8, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Herpes simplex virus-1 pUL56 degrades GOPC to alter the 2 plasma membrane proteome 3 4 1, 2Timothy K. Soh 5 1, 3Colin T. R. Davies 6 1, 2Julia Muenzner 7 2Viv Connor 8 2Clément R. Bouton 9 2Henry G. Barrow 10 2Cameron Smith 11 2Edward Emmott 12 3Robin Antrobus 13 2,4Stephen C. Graham 14 3,4Michael P. Weekes 15 *2,4,5Colin M. Crump 16 17 1These authors contributed equally 18 2Division of Virology, Department of Pathology, Cambridge University, Cambridge, CB2 19 1QP, UK 20 3Cambridge Institute for Medical Research, Cambridge University, Cambridge, CB2 21 0XY, UK 22 4Senior author 23 5Lead Contact. 24 *Correspondence: [email protected] 25 26 27 Keywords: Herpesvirus, Virus host interaction, Immune evasion, Membrane trafficking, 28 Proteasomal degradation, Quantitative proteomics, Uncharacterized ORF 29 1 bioRxiv preprint doi: https://doi.org/10.1101/729343; this version posted August 8, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 30 Summary 31 Herpesviruses are ubiquitous in the human population and they extensively remodel the 32 cellular environment during infection.
    [Show full text]
  • Cellular Immunology Regulation of Immune Responses by E3 Ubiquitin
    Cellular Immunology 340 (2019) 103878 Contents lists available at ScienceDirect Cellular Immunology journal homepage: www.elsevier.com/locate/ycimm Review article Regulation of immune responses by E3 ubiquitin ligase Cbl-b T ⁎ Rong Tanga, Wallace Y. Langdonb, Jian Zhangc, a Department of Nephrology, Xiangya Hospital, Central South University, Changsha, Hunan, PR China b School of Biological Sciences, University of Western Australia, Perth, Australia c Department of Pathology, The University of Iowa, Iowa City, IA, USA ARTICLE INFO ABSTRACT Keywords: Casitas B lymphoma-b (Cbl-b), a RING finger E3 ubiquitin ligase, has been identified as a critical regulator of Cbl-b adaptive immune responses. Cbl-b is essential for establishing the threshold for T cell activation and regulating Ubiquitination peripheral T cell tolerance through various mechanisms. Intriguingly, recent studies indicate that Cbl-b also Innate and adaptive immune responses modulates innate immune responses, and plays a key role in host defense to pathogens and anti-tumor immunity. T cell tolerance These studies suggest that targeting Cbl-b may represent a potential therapeutic strategy for the management of Immune-related disorders human immune-related disorders such as autoimmune diseases, infections, tumors, and allergic airway in- flammation. In this review, we summarize the latest developments regarding the roles of Cbl-b ininnateand adaptive immunity, and immune-mediated diseases. 1. Introduction adaptive immunity, and the involvement of Cbl-b in immune-mediated diseases. Ubiquitination, the covalent conjugation of ubiquitin (Ub) (a 76 amino-acid peptide) to protein substrates, is an essential mechanism of 2. The structures of Cbl family proteins post-translational modification, which modulates various cellular pathways.
    [Show full text]
  • NEDD4 Regulates Ubiquitination and Stability of the Cell Adhesion
    bioRxiv preprint doi: https://doi.org/10.1101/2021.02.07.430113; this version posted February 7, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. NEDD4 Regulates Ubiquitination and Stability of the Cell adhesion Molecule IGPR-1 via Lysosomal Pathway Linzi Sun1, Razie Amraei1, Nader Rahimi1* Department of Pathology, School of Medicine, Boston University Medical Campus, Boston, MA 02118, USA. Running title: NEDD4 targets IGPR-1 for ubiquitination and lysosomal degradation *Corresponding Author: Nader Rahimi, Departments of Pathology, Boston University Medical Campus, 670 Albany St., MA 02118, E-mail: [email protected]. Key words: NEDD4, IGPR-1, TMIGD2, CD28H, ubiquitination, lysosomal degradation Funding and additional information: This work was supported in part through grants from CTSI grant (UL1TR001430) and Malory Fund, Department of Pathology, Boston University. Data availability: Data and reagents are available from the corresponding author upon request. Conflicts of Interest: The authors declare no conflicts of interest. 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.02.07.430113; this version posted February 7, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. ABSTRACT: The cell adhesion molecule immunoglobulin and proline-rich receptor-1 (IGPR-1) regulates various critical cellular processes including, cell-cell adhesion, mechanosensing and autophagy.
    [Show full text]
  • Identification of a Herpes Simplex Virus 1 Gene Encoding Neurovirulence Factor by Chemical Proteomics
    ARTICLE https://doi.org/10.1038/s41467-020-18718-9 OPEN Identification of a herpes simplex virus 1 gene encoding neurovirulence factor by chemical proteomics Akihisa Kato1,2,3,8, Shungo Adachi 4,8, Shuichi Kawano 5, Kousuke Takeshima1, Mizuki Watanabe1, Shinobu Kitazume 6, Ryota Sato7, Hideo Kusano4, Naoto Koyanagi1,2,3, Yuhei Maruzuru1,2, Jun Arii1,2,3, ✉ ✉ Tomohisa Hatta4, Tohru Natsume 4 & Yasushi Kawaguchi 1,2,3 1234567890():,; Identification of the complete set of translated genes of viruses is important to understand viral replication and pathogenesis as well as for therapeutic approaches to control viral infection. Here, we use chemical proteomics, integrating bio-orthogonal non-canonical amino acid tagging and high-resolution mass spectrometry, to characterize the newly synthesized herpes simplex virus 1 (HSV-1) proteome in infected cells. In these infected cells, host cellular protein synthesis is shut-off, increasing the chance to preferentially detect viral proteomes. We identify nine previously cryptic orphan protein coding sequences whose translated products are expressed in HSV-1-infected cells. Functional characterization of one identified protein, designated piUL49, shows that it is critical for HSV-1 neurovirulence in vivo by regulating the activity of virally encoded dUTPase, a key enzyme that maintains accurate DNA replication. Our results demonstrate that cryptic orphan protein coding genes of HSV-1, and probably other large DNA viruses, remain to be identified. 1 Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan. 2 Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
    [Show full text]
  • The Ubiquitin Ligase Nedd4-1 Is Dispensable for the Regulation of PTEN Stability and Localization
    The ubiquitin ligase Nedd4-1 is dispensable for the regulation of PTEN stability and localization Fatemeh Fouladkou†‡, Tamara Landry‡§, Hiroshi Kawabe¶, Antje Neeb¶, Chen Lu†, Nils Brose¶, Vuk Stambolic§ʈ, and Daniela Rotin†ʈ †Hospital for Sick Children and Biochemistry Department, University of Toronto, MaRS-TMDT, 101 College Street, Toronto, Ontario M5G 1L7, Canada; §Ontario Cancer Institute, University Health Network, 610 University Avenue, and Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada; and ¶Department of Molecular Neurobiology, Max Planck Institute for Experimental Medicine, Go¨ttingen 37075, Germany Communicated by Aaron J. Ciechanover, Technion-Israel Institute of Technology, Bat Galim, Haifa, Israel, April 4, 2008 (received for review December 5, 2007) PTEN is a tumor suppressor frequently mutated in cancer. Recent location of this tumor suppressor to the nucleus, where it was reports implicated Nedd4-1 as the E3 ubiquitin ligase for PTEN that protected from degradation (22). regulates its stability and nuclear localization. We tested the Nedd4-1 is an E3 ubiquitin ligase that contains a C2 domain, physiological role of Nedd4-1 as a PTEN regulator by using cells and three or four WW domains, and a ubiquitin ligase Hect domain, and tissues derived from two independently generated strains of mice it belongs to the Nedd4 family of Hect E3 ligases that also includes with their Nedd4-1 gene disrupted. PTEN stability and ubiquitina- its relative Nedd4-2 (23–25). The WW domain of Nedd4 proteins tion were indistinguishable between the wild-type and Nedd4-1- recognizes and binds a short amino acid sequence in substrate deficient cells, and an interaction between the two proteins could proteins, called the PY motif (L/PPxY) (26–30).
    [Show full text]
  • E3 Ubiquitin Ligase NEDD4 Family‑Regulatory Network In
    Int. J. Biol. Sci. 2020, Vol. 16 2727 Ivyspring International Publisher International Journal of Biological Sciences 2020; 16(14): 2727-2740. doi: 10.7150/ijbs.48437 Review E3 Ubiquitin ligase NEDD4 family‑regulatory network in cardiovascular disease Ying Zhang1, Hao Qian1, Boquan Wu1, Shilong You1, Shaojun Wu1, Saien Lu1, Pingyuan Wang2, Liu Cao3, Naijin Zhang1 and Yingxian Sun1 1. Department of Cardiology, the First Hospital of China Medical University, Shenyang, Liaoning, P.R. China. 2. Staff scientist, Center for Molecular Medicine National Heart Lung and Blood Institute, National Institutes of Health, the United States. 3. Key Laboratory of Medical Cell Biology, Ministry of Education; Institute of Translational Medicine, China Medical University; Liaoning Province Collaborative Innovation Center of Aging Related Disease Diagnosis and Treatment and Prevention, Shenyang, Liaoning, China. Corresponding authors: 155 Nanjing North Street, Heping District, Shenyang, 110001, Liaoning Province, People’s Republic of China. 77 Puhe Road, Shenbei New District, Shenyang, 110001, Liaoning Province, People’s Republic of China. Telephone Number: +86 15040171605; +86 13804068889; +86 18900911888, E-mail: [email protected] (Y.S); [email protected] (N.Z); [email protected] (L.C). © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. Received: 2020.05.20; Accepted: 2020.08.06; Published: 2020.08.21 Abstract Protein ubiquitination represents a critical modification occurring after translation. E3 ligase catalyzes the covalent binding of ubiquitin to the protein substrate, which could be degraded.
    [Show full text]