DOCSLIB.ORG

Characterisation of Serpinb2 As a Stress Response Modulator

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

University of Wollongong Research Online University of Wollongong Thesis Collection 1954-2016 University of Wollongong Thesis Collections 2015 Characterisation of SerpinB2 as a stress response modulator Jodi Anne Lee University of Wollongong Follow this and additional works at: https://ro.uow.edu.au/theses University of Wollongong Copyright Warning You may print or download ONE copy of this document for the purpose of your own research or study. The University does not authorise you to copy, communicate or otherwise make available electronically to any other person any copyright material contained on this site. You are reminded of the following: This work is copyright. Apart from any use permitted under the Copyright Act 1968, no part of this work may be reproduced by any process, nor may any other exclusive right be exercised, without the permission of the author. Copyright owners are entitled to take legal action against persons who infringe their copyright. A reproduction of material that is protected by copyright may be a copyright infringement. A court may impose penalties and award damages in relation to offences and infringements relating to copyright material. Higher penalties may apply, and higher damages may be awarded, for offences and infringements involving the conversion of material into digital or electronic form. Unless otherwise indicated, the views expressed in this thesis are those of the author and do not necessarily represent the views of the University of Wollongong. Recommended Citation Lee, Jodi Anne, Characterisation of SerpinB2 as a stress response modulator, Doctor of Philosophy thesis, School of Biological Sciences, University of Wollongong, 2015. https://ro.uow.edu.au/theses/4538 Research Online is the open access institutional repository for the University of Wollongong. For further information contact the UOW Library: [email protected] Characterisation of SerpinB2 as a stress response modulator A thesis submitted in fulfilment of the requirements for the award of the degree Doctor of Philosophy from University of Wollongong By Jodi Anne Lee Bachelor of Science (Honours 1) School of Biological Sciences University of Wollongong 2015 I Declaration This thesis is submitted in accordance with the regulations of the University of Wollongong in fulfilment of the higher degree in research of Doctorate of Philosophy. It does not contain any material previously published by another person except where due reference is made in the text. The experimental work described in this thesis is original and has not been submitted for a degree at any other University. Jodi Anne Lee - II - List of Publications Lee JA, Yerbury JJ, Farrawell N, Shearer RF, Constantinescu P, Hatters DM, Schroder WA, Suhrbier A, Wilson MR, Saunders DN, Ranson, M. SerpinB2 (PAI- 2) Modulates Proteostasis via Binding Misfolded Proteins and Promotion of Cytoprotective Inclusion Formation. PLoS One 2015; 10(6). Epub 2015 June 18. PMC4470917. Yerbury JJ, Gower D, Vanags L, Roberts K, Lee JA, Ecroyd H. The small heat shock proteins αB-crystallin and Hsp27 suppress SOD1 aggregation in vitro. Cell Stress Chaperones 2013 Mar; 18(2):251-7. Epub 2012 Sep 21. PMID: 22993064 PMCID: PMC3581626 Vine KL, Chandran VI, Locke JM, Matesic L, Lee J, Skropeta D, Bremner JB, Ranson M. Targeting urokinase and the transferrin receptor with novel, anti-mitotic N-alkylisatin cytotoxin conjugates causes selective cancer cell death and reduces tumor growth. Curr Cancer Drug Targets. 2012 Jan 1;12(1):64-73. PubMed PMID: 22111834. Lee JA, Cochran BJ, Lobov S, Ranson M. Forty years later and the role of plasminogen activator inhibitor type 2/SERPINB2 is still an enigma. Semin Thromb Hemost. 2011 Jun;37(4):395-407. Epub 2011 Jul 30. Review. PubMed PMID: 21805446. De Souza M, Matthews H, Lee JA, Ranson M, Kelso MJ. Small molecule antagonists of the urokinase (uPA): urokinase receptor (uPAR) interaction with high reported potencies show only weak effects in cell-based competition assays employing the native uPAR ligand. Bioorg Med Chem. 2011 Apr 15;19(8):2549-56. Epub 2011 Mar 12. PubMed PMID: 21454081. Lee JA, Croucher DR, Ranson M. Differential endocytosis of tissue plasminogen activator by serpins SerpinE1 and SerpinB2 on human peripheral blood monocytes. Thromb Haemost. 2010 Dec;104(6):1133-42. Epub 2010 Sep 13. PubMed PMID: 20838737. Cochran BJ, Gunawardhana LP, Vine KL, Lee JA, Lobov S, Ranson M. The CD- loop of SerpinB2 (SERPINB2) is redundant in the targeting, inhibition and clearance of cell surface uPA activity. BMC Biotechnol. 2009 May 14;9:43. PubMed PMID:19442270; PubMed Central PMCID: PMC2693429. - III - List of Presentations Poster Vine KL, Chandran VI, Locke JM, Matesic L, Lee J, Skropeta D, Bremner JB, and Ranson M. Targeting Urokinase and the Transferrin Receptor with Novel, Anti- Mitotic N-Alkylisatin Cytotoxin Conjugates Causes Selective Cancer Cell Death and Reduces Tumour Growth. 24th Lorne Cancer Conference, Lorne Victoria, Feb. 2012 Lee JA, Croucher DR and Ranson M. Differential endocytosis of Tissue Plasminogen Activator by Serpins PAI-1 and PAI-2 on human peripheral blood monocytes. Australian Society for Medical Research (NSW) Scientific Meeting. The Powerhouse Museum, Ultimo. (2010) Vine KL, Locke JM, Lee JA, Bremner, JB, and Ranson M. Synthesis and in vitro anticancer activity of a targeted plasminogen activator inhibitor type II (PAI-2) cytotoxin conjugate. 7th International Symposium On Targeted Anticancer Therapies, Amsterdam, The Netherlands, March 2009 Lee JA, Croucher DR and Ranson M The Plasminogen Activation System in Monocytic Cell Differentiation and Proliferation: potential target for plasminogen activation inhibitor type 2 based therapeutics. XIth International Workshop on Molecular & Cellular Biology of Plasminogen Activation, Saltsjöbaden, Sweden. (2007) Oral Lee JA, Saunders DN, Schroder W, Wilson M, Yerbury JJ, Marie Ranson. SerpinB2 (PAI-2) modulates protective cellular responses to proteotoxic stress and processing of intracellular protein aggregates. Plasminogen Activation System in Pathology Workshop, Illawarra Medical Research Institute, University of Wollongong NSW Australia. (2012) Lee JA, Croucher DR and Ranson M Cell surface interactions of Tissue Plasminogen Activator with peripheral blood monocytes. XIIth International Workshop on Molecular & Cellular Biology of Plasminogen Activation, Cold Spring Habor Laboratory, Cold Spring Harbor, New York. (2009) - IV - Acknowledgements I would like to sincerely thank Professor Marie Ranson, Dr Justin Yerbury and Dr David Croucher (Chapters 2 and 3) for the formal supervision of the work within this thesis. I will be forever grateful for the contributions they have made to the projects undertaken and for all that they have taught me. I would like to acknowledge and thank Dr Darren Saunders, Professor Mark Wilson, Dr Andrew Aquilina and Associate Professor Heath Ecroyd for the many lengthy discussions on proteastasis networks and experimental advice given, without which I would not have gotten so far. I would also like to thank Dr Wayne Schroder for supporting the working hypothesis for a role of SerpinB2 as a molecular chaperone and providing the mouse embryonic fibroblasts used within the latter part of this thesis, in which to test it. I will be forever grateful to Professor Robert Medcalf for believing me when I approached him in New York to discuss this work, in its infancy. I would like to thank all fellow PhD students whom I have worked alongside for the contribution they made to my health and wellbeing. I wish you all the best science can offer. - V - Abstract SerpinB2 [plasminogen activator inhibitor type-2 (PAI-2)] was first described as a placental protein, which regulated plasminogen activation via the inhibition of the urokinase (uPA) or tissue-type (tPA) plasminogen activators during pregnancy. Further work has confirmed that SerpinB2 is an efficient inhibitor of cell surface, receptor bound uPA and clearance of the serpin:protease complex occurs via members of the low density lipoprotein receptor (LDLR) family, specifically very low density lipoprotein receptor (VLDLR) and low density lipoprotein-related protein receptor (LRP). This property has allowed for the development of SerpinB2 drug conjugates for the selective targeting of uPA positive carcinoma cells. However, potential utility of SerpinB2 in haematological disorders, such as acute myeloid leukaemia (AML), as well as its role as a physiological inhibitor of tPA is not well characterised. To date biochemical analysis has shown that SerpinB2 is able to inhibit the two-chain (active) form of tPA. However, whether this inhibitory function occurs within the cellular context was unknown. This thesis investigates the potential use of exogenous SerpinB2 as a delivery vehicle for the specific targeting of lymphoma or leukaemia, via cell surface uPA using the relatively low versus high uPA expressing, monocytic cell lines U-937 and THP-1, respectively, as disease models. We show here that uPA:SerpinB2 is internalised via the Low Density Lipoprotein Receptor (LDLR) family members, specifically Low Density Lipoprotein-Related Protein (LRP) and Very Low Density - VI - Lipoprotein Receptor (VLDLR) in a uPA dependent manner and these findings are consistent with those previously reported for epithelial cancer cell lines. In order to clarify the role of SerpinB2 in the inhibition and clearance of tPA from the cell surface of monocytes and macrophages, we next assessed the formation and endocytosis of tPA:SerpinB2
Recommended publications
  • Serpinb1 Controls Encephalitogenic T Helper Cells in Neuroinflammation

    Serpinb1 Controls Encephalitogenic T Helper Cells in Neuroinflammation

    SerpinB1 controls encephalitogenic T helper cells in neuroinflammation Lifei Houa,b,1,2, Deepak A. Raoc,d, Koichi Yukie,f, Jessica Cooleya, Lauren A. Hendersonb,g, A. Helena Jonssonc,d, Dion Kaisermanh, Mark P. Gormanb,i, Peter A. Nigrovicc,d,g, Phillip I. Birdh, Burkhard Becherj, and Eileen Remold-O’Donnella,b,k,2 aThe Program in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA 02115; bDepartment of Pediatrics, Harvard Medical School, Boston, MA 02115; cDivision of Rheumatology, Immunology and Allergy, Brigham and Women’s Hospital, Boston, MA 02115; dDepartment of Medicine, Harvard Medical School, Boston, MA 02115; eDepartment of Anesthesiology, Critical Care and Pain Medicine, Boston Children’s Hospital, Boston, MA 02115; fDepartment of Anesthesiology, Harvard Medical School, Boston, MA 02115; gDivision of Immunology, Boston Children’s Hospital, Boston, MA 02115; hDepartment of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Melbourne, VIC, 3800, Australia; iDepartment of Neurology, Boston Children’s Hospital, Boston, MA 02115; jInflammation Unit, Institute of Experimental Immunology, University of Zurich, CH-8057 Zurich, Switzerland; and kDepartment of Hematology/Oncology, Harvard Medical School, Boston, MA 02115 Edited by Jean Laurent-Casanova, Rockefeller University, New York, NY, and approved August 27, 2019 (received for review April 5, 2019) SerpinB1, a protease inhibitor and neutrophil survival factor, was flammatory tissue injury and neutrophil death, and in naïve mice, recently linked with IL-17–expressing T cells. Here, we show that preserves the bone marrow reserve of mature neutrophils by serpinB1 (Sb1) is dramatically inducedinasubsetofeffector restricting spontaneous cell death mediated by the granule serine CD4 cells in experimental autoimmune encephalomyelitis (EAE).
  • ONCOGENOMICS Through the Use of Chemotherapeutic Agents

    ONCOGENOMICS Through the Use of Chemotherapeutic Agents

    Oncogene (2002) 21, 7749 – 7763 ª 2002 Nature Publishing Group All rights reserved 0950 – 9232/02 $25.00 www.nature.com/onc Expression profiling of primary non-small cell lung cancer for target identification Jim Heighway*,1, Teresa Knapp1, Lenetta Boyce1, Shelley Brennand1, John K Field1, Daniel C Betticher2, Daniel Ratschiller2, Mathias Gugger2, Michael Donovan3, Amy Lasek3 and Paula Rickert*,3 1Target Identification Group, Roy Castle International Centre for Lung Cancer Research, University of Liverpool, 200 London Road, Liverpool L3 9TA, UK; 2Institute of Medical Oncology, University of Bern, 3010 Bern, Switzerland; 3Incyte Genomics Inc., 3160 Porter Drive, Palo Alto, California, CA 94304, USA Using a panel of cDNA microarrays comprising 47 650 Introduction transcript elements, we have carried out a dual-channel analysis of gene expression in 39 resected primary The lack of generally effective therapies for lung cancer human non-small cell lung tumours versus normal lung leads to the high mortality rate associated with this tissue. Whilst *11 000 elements were scored as common disease. Recorded 5-year survival figures vary, differentially expressed at least twofold in at least one to some extent perhaps reflecting differences in sample, 96 transcripts were scored as over-represented ascertainment methods, but across Europe are approxi- fourfold or more in at least seven out of 39 tumours mately 10% (Bray et al., 2002). Whilst surgery is an and 30 sequences 16-fold in at least two out of 39 effective strategy for the treatment of localized lesions, tumours, including 24 transcripts in common. Tran- most patients present with overtly or covertly dissemi- scripts (178) were found under-represented fourfold in nated disease.
  • Biomarkers of Neonatal Skin Barrier Adaptation Reveal Substantial Differences Compared to Adult Skin

    Biomarkers of Neonatal Skin Barrier Adaptation Reveal Substantial Differences Compared to Adult Skin

    www.nature.com/pr CLINICAL RESEARCH ARTICLE OPEN Biomarkers of neonatal skin barrier adaptation reveal substantial differences compared to adult skin Marty O. Visscher1,2, Andrew N. Carr3, Jason Winget3, Thomas Huggins3, Charles C. Bascom3, Robert Isfort3, Karen Lammers1 and Vivek Narendran1 BACKGROUND: The objective of this study was to measure skin characteristics in premature (PT), late preterm (LPT), and full-term (FT) neonates compared with adults at two times (T1, T2). METHODS: Skin samples of 61 neonates and 34 adults were analyzed for protein biomarkers, natural moisturizing factor (NMF), and biophysical parameters. Infant groups were: <34 weeks (PT), 34–<37 weeks (LPT), and ≥37 weeks (FT). RESULTS: Forty proteins were differentially expressed in FT infant skin, 38 in LPT infant skin, and 12 in PT infant skin compared with adult skin at T1. At T2, 40 proteins were differentially expressed in FT infants, 38 in LPT infants, and 54 in PT infants compared with adults. All proteins were increased at both times, except TMG3, S100A7, and PEBP1, and decreased in PTs at T1. The proteins are involved in filaggrin processing, protease inhibition/enzyme regulation, and antimicrobial function. Eight proteins were decreased in PT skin compared with FT skin at T1. LPT and FT proteins were generally comparable at both times. Total NMF was lower in infants than adults at T1, but higher in infants at T2. CONCLUSIONS: Neonates respond to the physiological transitions at birth by upregulating processes that drive the production of lower pH of the skin and water-binding NMF components, prevent protease activity leading to desquamation, and increase the 1234567890();,: barrier antimicrobial properties.
  • Propranolol-Mediated Attenuation of MMP-9 Excretion in Infants with Hemangiomas

    Propranolol-Mediated Attenuation of MMP-9 Excretion in Infants with Hemangiomas

    Supplementary Online Content Thaivalappil S, Bauman N, Saieg A, Movius E, Brown KJ, Preciado D. Propranolol-mediated attenuation of MMP-9 excretion in infants with hemangiomas. JAMA Otolaryngol Head Neck Surg. doi:10.1001/jamaoto.2013.4773 eTable. List of All of the Proteins Identified by Proteomics This supplementary material has been provided by the authors to give readers additional information about their work. © 2013 American Medical Association. All rights reserved. Downloaded From: https://jamanetwork.com/ on 10/01/2021 eTable. List of All of the Proteins Identified by Proteomics Protein Name Prop 12 mo/4 Pred 12 mo/4 Δ Prop to Pred mo mo Myeloperoxidase OS=Homo sapiens GN=MPO 26.00 143.00 ‐117.00 Lactotransferrin OS=Homo sapiens GN=LTF 114.00 205.50 ‐91.50 Matrix metalloproteinase‐9 OS=Homo sapiens GN=MMP9 5.00 36.00 ‐31.00 Neutrophil elastase OS=Homo sapiens GN=ELANE 24.00 48.00 ‐24.00 Bleomycin hydrolase OS=Homo sapiens GN=BLMH 3.00 25.00 ‐22.00 CAP7_HUMAN Azurocidin OS=Homo sapiens GN=AZU1 PE=1 SV=3 4.00 26.00 ‐22.00 S10A8_HUMAN Protein S100‐A8 OS=Homo sapiens GN=S100A8 PE=1 14.67 30.50 ‐15.83 SV=1 IL1F9_HUMAN Interleukin‐1 family member 9 OS=Homo sapiens 1.00 15.00 ‐14.00 GN=IL1F9 PE=1 SV=1 MUC5B_HUMAN Mucin‐5B OS=Homo sapiens GN=MUC5B PE=1 SV=3 2.00 14.00 ‐12.00 MUC4_HUMAN Mucin‐4 OS=Homo sapiens GN=MUC4 PE=1 SV=3 1.00 12.00 ‐11.00 HRG_HUMAN Histidine‐rich glycoprotein OS=Homo sapiens GN=HRG 1.00 12.00 ‐11.00 PE=1 SV=1 TKT_HUMAN Transketolase OS=Homo sapiens GN=TKT PE=1 SV=3 17.00 28.00 ‐11.00 CATG_HUMAN Cathepsin G OS=Homo
  • A Truncating Mutation in SERPINB6 Is Associated with Autosomal-Recessive Nonsyndromic Sensorineural Hearing Loss

    A Truncating Mutation in SERPINB6 Is Associated with Autosomal-Recessive Nonsyndromic Sensorineural Hearing Loss

    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector REPORT A Truncating Mutation in SERPINB6 Is Associated with Autosomal-Recessive Nonsyndromic Sensorineural Hearing Loss Aslı Sırmacı,1,2 Seyra Erbek,3 Justin Price,1,2 Mingqian Huang,4 Duygu Duman,5 F. Basxak Cengiz,5 Gu¨ney Bademci,1,2 Suna Tokgo¨z-Yılmaz,5 Burcu Hisxmi,5 Hilal O¨ zdag˘,6 Banu O¨ ztu¨rk,7 Sevsen Kulaksızog˘lu,8 Erkan Yıldırım,9 Haris Kokotas,10 Maria Grigoriadou,10 Michael B. Petersen,10 Hashem Shahin,11 Moien Kanaan,11 Mary-Claire King,12 Zheng-Yi Chen,4 Susan H. Blanton,1,2 Xue Z. Liu,2,13 Stephan Zuchner,1,2 Nejat Akar,5 and Mustafa Tekin1,2,* More than 270 million people worldwide have hearing loss that affects normal communication. Although astonishing progress has been made in the identification of more than 50 genes for deafness during the past decade, the majority of deafness genes are yet to be identified. In this study, we mapped a previously unknown autosomal-recessive nonsyndromic sensorineural hearing loss locus (DFNB91) to chromo- some 6p25 in a consanguineous Turkish family. The degree of hearing loss was moderate to severe in affected individuals. We subsequently identified a nonsense mutation (p.E245X) in SERPINB6, which is located within the linkage interval for DFNB91 and encodes for an intra- cellular protease inhibitor. The p.E245X mutation cosegregated in the family as a completely penetrant autosomal-recessive trait and was absent in 300 Turkish controls. The mRNA expression of SERPINB6 was reduced and production of protein was absent in the peripheral leukocytes of homozygotes, suggesting that the hearing loss is due to loss of function of SERPINB6.
  • Epigenetic Suppression of SERPINB1 Promotes Inflammation-Mediated Prostate Cancer Progression

    Epigenetic Suppression of SERPINB1 Promotes Inflammation-Mediated Prostate Cancer Progression

    Published OnlineFirst January 4, 2019; DOI: 10.1158/1541-7786.MCR-18-0638 Cancer Genes and Networks Molecular Cancer Research Epigenetic Suppression of SERPINB1 Promotes Inflammation-Mediated Prostate Cancer Progression Irina Lerman1, Xiaoting Ma1, Christina Seger1, Aerken Maolake2, Maria de la Luz Garcia-Hernandez3, Javier Rangel-Moreno3, Jessica Ackerman4, Kent L. Nastiuk2, Martha Susiarjo5, and Stephen R. Hammes1 Abstract Granulocytic myeloid infiltration and resultant enhanced tatic epithelial cells (RWPE-1) increases proliferation, neutrophil elastase (NE) activity is associated with poor out- decreases apoptosis, and stimulates expression of epithelial- comes in numerous malignancies. We recently showed that NE to-mesenchymal transition markers. In contrast, stable SER- expression and activity from infiltrating myeloid cells was high PINB1 expression in normally low-expressing prostate cancer in human prostate cancer xenografts and mouse Pten-null cells (C4-2) reduces xenograft growth in vivo. Finally, EZH2- prostate tumors. We further demonstrated that NE directly mediated histone (H3K27me3) methylation and DNA stimulated human prostate cancer cells to proliferate, migrate, methyltransferase–mediated DNA methylation suppress and invade, and inhibition of NE in vivo attenuated xenograft SERPINB1 expression in prostate cancer cells. Analysis of growth. Interestingly, reduced expression of SERPINB1, an The Cancer Genome Atlas and pyrosequencing demonstrate endogenous NE inhibitor, also correlates with diminished hypermethylation of the SERPINB1 promoter in prostate survival in some cancers. Therefore, we sought to characterize cancer compared with normal tissue, and the extent of pro- the role of SERPINB1 in prostate cancer. We find that SER- moter methylation negatively correlates with SERPINB1 PINB1 expression is reduced in human metastatic and locally mRNA expression.
  • Pediatric Primitive Neuroectodermal Tumors of the Central Nervous System Differentially Express Granzyme Inhibitors

    Pediatric Primitive Neuroectodermal Tumors of the Central Nervous System Differentially Express Granzyme Inhibitors

    RESEARCH ARTICLE Pediatric Primitive Neuroectodermal Tumors of the Central Nervous System Differentially Express Granzyme Inhibitors Jeroen F. Vermeulen1, Wim van Hecke1, Wim G. M. Spliet1, José Villacorta Hidalgo3, Paul Fisch3, Roel Broekhuizen1, Niels Bovenschen1,2* 1 Department of Pathology, University Medical Center Utrecht, 3584CX, Utrecht, The Netherlands, 2 Laboratory of Translational Immunology, University Medical Center Utrecht, 3584CX, Utrecht, The Netherlands, 3 Institute of Pathology, University Medical Center Freiburg, 79106, Freiburg, Germany * [email protected] Abstract Background OPEN ACCESS Central nervous system (CNS) primitive neuroectodermal tumors (PNETs) are malignant Citation: Vermeulen JF, van Hecke W, Spliet WGM, primary brain tumors that occur in young infants. Using current standard therapy, up to 80% Villacorta Hidalgo J, Fisch P, Broekhuizen R, et al. of the children still dies from recurrent disease. Cellular immunotherapy might be key to (2016) Pediatric Primitive Neuroectodermal Tumors improve overall survival. To achieve efficient killing of tumor cells, however, immunotherapy of the Central Nervous System Differentially Express Granzyme Inhibitors. PLoS ONE 11(3): e0151465. has to overcome cancer-associated strategies to evade the cytotoxic immune response. doi:10.1371/journal.pone.0151465 Whether CNS-PNETs can evade the immune response remains unknown. Editor: Javier S Castresana, University of Navarra, SPAIN Methods Received: September 3, 2015 We examined by immunohistochemistry the immune response and immune evasion strate- Accepted: February 29, 2016 gies in pediatric CNS-PNETs. Published: March 10, 2016 Copyright: © 2016 Vermeulen et al. This is an open Results access article distributed under the terms of the Creative Commons Attribution License, which permits Here, we show that CD4+, CD8+, γδ-T-cells, and Tregs can infiltrate pediatric CNS-PNETs, unrestricted use, distribution, and reproduction in any although the activation status of cytotoxic cells is variable.
  • Serpinb2 Is Involved in Cellular Response Upon UV Irradiation

    Serpinb2 Is Involved in Cellular Response Upon UV Irradiation

    www.nature.com/scientificreports OPEN SerpinB2 is involved in cellular response upon UV irradiation Hajnalka Majoros1, Zsuzsanna Ujfaludi1, Barbara Nikolett Borsos1, Viktória Vivien Hudacsek1, Zita Nagy3, Frederic Coin3, Krisztina Buzas2, Ilona Kovács4, Tamás Bíró5,6, Imre Miklós Boros1,2 1 Received: 20 August 2018 & Tibor Pankotai Accepted: 10 December 2018 Ultraviolet light induced pyrimidine dimer is a helix distortion DNA damage type, which recruits repair Published: xx xx xxxx complexes. However, proteins of these complexes that take part in both DNA damage recognition and repair have been well-described, the regulation of the downstream steps of nucleotide excision repair (NER) have not been clearly clarifed yet. In a high-throughput screen, we identifed SerpinB2 (SPB2) as one of the most dramatically upregulated gene in keratinocytes following UV irradiation. We found that both the mRNA and the protein levels of SPB2 were increased upon UV irradiation in various cell lines. Additionally, UV damage induced translocation of SPB2 from the cytoplasm to the nucleus as well as the damage induced foci formation of it. Here we show that SPB2 co-localizes with XPB involved in the NER pathway at UV-induced repair foci. Finally, we demonstrated that UV irradiation promoted the association of SPB2 with ubiquitylated proteins. In basal cell carcinoma tumour cells, we identifed changes in the subcellular localization of SPB2. Based on our results, we conclude that SPB2 protein has a novel role in UV-induced NER pathway, since it regulates the removal of the repair complex from the damaged site leading to cancerous malformation. Our genome is constantly exposed to endogenous and exogenous sources leading to DNA damage and impair- ment of genome integrity.
  • Differential Gene Expression of Serine Protease Inhibitors in Bovine

    Differential Gene Expression of Serine Protease Inhibitors in Bovine

    Hayashi et al. Reproductive Biology and Endocrinology 2011, 9:72 http://www.rbej.com/content/9/1/72 RESEARCH Open Access Differential gene expression of serine protease inhibitors in bovine ovarian follicle: possible involvement in follicular growth and atresia Ken-Go Hayashi, Koichi Ushizawa, Misa Hosoe and Toru Takahashi* Abstract Background: SERPINs (serine protease inhibitors) regulate proteases involving fibrinolysis, coagulation, inflammation, cell mobility, cellular differentiation and apoptosis. This study aimed to investigate differentially expressed genes of members of the SERPIN superfamily between healthy and atretic follicles using a combination of microarray and quantitative real-time PCR (QPCR) analysis. In addition, we further determined mRNA and protein localization of identified SERPINs in estradiol (E2)-active and E2-inactive follicles by in situ hybridization and immunohistochemistry. Methods: We performed microarray analysis of healthy (10.7 +/- 0.7 mm) and atretic (7.8 +/- 0.2 mm) follicles using a custom-made bovine oligonucleotide microarray to screen differentially expressed genes encoding SERPIN superfamily members between groups. The expression profiles of six identified SERPIN genes were further confirmed by QPCR analysis. In addition, mRNA and protein localization of four SERPINs was investigated in E2- active and E2-inactive follicles using in situ hybridization and immunohistochemistry. Results: We have identified 11 SERPIN genes expressed in healthy and atretic follicles by microarray analysis. QPCR analysis confirmed that mRNA expression of four SERPINs (SERPINA5, SERPINB6, SERPINE2 and SERPINF2) was greater in healthy than in atretic follicles, while two SERPINs (SERPINE1 and SERPING1) had greater expression in atretic than in healthy follicles. In situ hybridization showed that SERPINA5, SERPINB6 and SERPINF2 mRNA were localized in GCs of E2-active follicles and weakly expressed in GCs of E2-inactive follicles.
  • Adaptive Evolution and Divergence of SERPINB3: a Young Duplicate in Great Apes

    Adaptive Evolution and Divergence of SERPINB3: a Young Duplicate in Great Apes

    Adaptive Evolution and Divergence of SERPINB3: A Young Duplicate in Great Apes Sı´lvia Gomes1*, Patrı´cia I. Marques1,2, Rune Matthiesen3, Susana Seixas1* 1 Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal, 2 Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal, 3 National Health Institute Doutor Ricardo Jorge (INSA), Lisboa, Portugal Abstract A series of duplication events led to an expansion of clade B Serine Protease Inhibitors (SERPIN), currently displaying a large repertoire of functions in vertebrates. Accordingly, the recent duplicates SERPINB3 and B4 located in human 18q21.3 SERPIN cluster control the activity of different cysteine and serine proteases, respectively. Here, we aim to assess SERPINB3 and B4 coevolution with their target proteases in order to understand the evolutionary forces shaping the accelerated divergence of these duplicates. Phylogenetic analysis of primate sequences placed the duplication event in a Hominoidae ancestor (,30 Mya) and the emergence of SERPINB3 in Homininae (,9 Mya). We detected evidence of strong positive selection throughout SERPINB4/B3 primate tree and target proteases, cathepsin L2 (CTSL2) and G (CTSG) and chymase (CMA1). Specifically, in the Homininae clade a perfect match was observed between the adaptive evolution of SERPINB3 and cathepsin S (CTSS) and most of sites under positive selection were located at the inhibitor/protease interface. Altogether our results seem to favour a coevolution hypothesis for SERPINB3, CTSS and CTSL2 and for SERPINB4 and CTSG and CMA1.A scenario of an accelerated evolution driven by host-pathogen interactions is also possible since SERPINB3/B4 are potent inhibitors of exogenous proteases, released by infectious agents.
  • Alpha -Antitrypsin Deficiency

    Alpha -Antitrypsin Deficiency

    The new england journal of medicine Review Article Dan L. Longo, M.D., Editor Alpha1-Antitrypsin Deficiency Pavel Strnad, M.D., Noel G. McElvaney, D.Sc., and David A. Lomas, Sc.D.​​ lpha1-antitrypsin (AAT) deficiency is one of the most common From the Department of Internal Med­ genetic diseases. Most persons carry two copies of the wild-type M allele icine III, University Hospital RWTH of SERPINA1, which encodes AAT, and have normal circulating levels of the (Rheinisch–Westfälisch Technische Hoch­ A schule) Aachen, Aachen, Germany (P.S.); protein. Ninety-five percent of severe cases of AAT deficiency result from the homo- the Irish Centre for Genetic Lung Dis­ zygous substitution of a single amino acid, Glu342Lys (the Z allele), which is present ease, Royal College of Surgeons in Ire­ in 1 in 25 persons of European descent (1 in 2000 persons of European descent land, Beaumont Hospital, Dublin (N.G.M.); and UCL Respiratory, Division of Medi­ are homozygotes). Mild AAT deficiency typically results from a different amino cine, Rayne Institute, University College acid replacement, Glu264Val (the S allele), which is found in 1 in 4 persons in the London, London (D.A.L.). Address re­ Iberian peninsula. However, many other alleles have been described that have vari- print requests to Dr. Lomas at UCL Re­ spiratory, Rayne Institute, University Col­ able effects, such as a lack of protein production (null alleles), production of mis- lege London, London WC1E 6JF, United folded protein, or no effect on the level or function of circulating AAT (Table 1). Kingdom, or at d .
  • Pso P27, a SERPINB3/B4-Derived Protein, Is Most Likely a Common Autoantigen in Chronic Inflammatory Diseases

    Pso P27, a SERPINB3/B4-Derived Protein, Is Most Likely a Common Autoantigen in Chronic Inflammatory Diseases

    Clinical Immunology 174 (2017) 10–17 Contents lists available at ScienceDirect Clinical Immunology journal homepage: www.elsevier.com/locate/yclim Pso p27, a SERPINB3/B4-derived protein, is most likely a common autoantigen in chronic inflammatory diseases Ole-Jan Iversen a,⁎,HildeLysvanda, Geir Slupphaug b a Department of Laboratory Medicine, Children's and Women's Health, Faculty of Medicine, Norwegian University of Science and Technology, NTNU, Trondheim, Norway b Department of Cancer Research and Molecular Medicine, Faculty of Medicine and PROMEC Core Facility for Proteomics and Metabolomics, Norwegian University of Science and Technology, NTNU, Trondheim, Norway article info abstract Article history: Autoimmune diseases are characterized by chronic inflammatory reactions localized to an organ or organ- Received 17 October 2016 system. They are caused by loss of immunologic tolerance toward self-antigens, causing formation of autoanti- Received in revised form 1 November 2016 bodies that mistakenly attack their own body. Psoriasis is a chronic inflammatory autoimmune skin disease in accepted with revision 13 November 2016 which the underlying molecular mechanisms remain elusive. In this review, we present evidence accumulated Available online 15 November 2016 through more than three decades that the serpin-derived protein Pso p27 is an autoantigen in psoriasis and prob- ably also in other chronic inflammatory diseases. Keywords: Autoimmune diseases Pso p27 is derived from the serpin molecules SERPINB3 and SERPINB4 through non-canonical cleavage by mast Pso p27 cell chymase. In psoriasis, it is exclusively found in skin lesions and not in uninvolved skin. The serpins are cleaved SERPINB3/B4 into three fragments that remain associated as a Pso p27 complex with novel immunogenic properties and in- Mast cells creased tendency to form large aggregates compared to native SERPINB3/B4.