DOCSLIB.ORG

Supporting Information

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Supporting Information Supporting Information McLaughlin et al. 10.1073/pnas.0909777106 Fig. S1. Growth of the most resistant 15 deletion strains compared to the wild-type. The wild-type and the most resistant 15 deletion strains were grown in liquid YPD media for 20 h at 30 °C on different concentrations of Tcin. McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 1of9 Fig. S2. Viability of BY4743 and the most resistant 15 mutants. Serial dilutions were spotted on YPD plates containing 12 ␮M Tcin (A)or24␮M Tcin (B). McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 2of9 Fig. S3. Growth of the most resistant 15 strains in the presence of Tcin in YPD or YPG media. The most resistant 15 strains were grown on 2, 4, or 8 ␮M Tcin in liquid YPD or YPG media. Cells were incubated at 30 °C for 20 h in YPD media or for 48 h at 30 °C in YPG media. Relative growth was calculated as the ratio between the growth of treated cells to that of untreated cells with a ratio of 1.0, indicating no effect on growth. McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 3of9 Fig. S4. Relative growth of BY4743 and the top 15 Tcin-resistant mutants in the presence of cycloheximide or hygromycin. (A) Cells were grown in 0.5 ␮g/mL or 1 ␮g/mL cycloheximide. (B) Cells were grown in 10 ␮g/mL hygromycin B in either liquid YPD or YPG media. Relative growth was calculated as the ratio between growth of treated cells to that of untreated cells with a ratio of 1.0, indicating no effect on growth. Error bars, the standard error of two independent experiments with each experiment performed in triplicate. McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 4of9 Fig. S5. Growth of the 15 Tcin resistant strains in the presence of anisomycin in YPD or YPG media. The 15 Tcin resistant strains were grown in YPD or YPG liquid media containing 2.5, 5, or 7.5 ␮g/mL anisomycin. Cells in YPD media were incubated at 30 °C for 20 h, while cells in YPG media were grown for 48 h at 30 °C. Relative growth was calculated as the ratio between growth of treated cells to that of untreated cells with a ratio of 1.0, indicating no effect on growth. McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 5of9 Fig. S6. Effects of Tcin on total and mitochondrial translation in the most resistant 15 deletion strains. Total translation (A) and mitochondrial translation (B) for the wild-type and the most resistant 15 deletion mutants is shown. Samples were treated with 8 ␮M Tcin/OD600 of cells for1hasdescribed in the Materials and Methods. Each graph represents the mean of four independent experiments with error bars representing the standard deviation. McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 6of9 Fig. S7. Verification of the top 15 Tcin-resistant deletion strains and the BY4743 rho0 strain. (A) Each strain was verified by PCR amplification using specific cassette primers unique to each strain as described in the Materials and Methods. The amplification products were separated on a 1% agarose gel. MW denotes the molecular marker. The parental strain, BY4743, is included as the negative control. (B) Southern blot analysis of the BY4743 strain (rhoϩ) and a rho0 strain derived from the parental BY4743 strain using COX1 as a probe. The lower panel shows the ethidium bromide stained DNA separated on a 1% agarose gel, indicating equal loading of the samples. McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 7of9 Table S1. S. cerevisiae strains that showed resistance to 4 ␮M Tcin* GO Process term Gene(s) Translation MRP21, FES1, CBP6, MRPL36, MRPS5, MRPL37, MRPL27, MRPL32, SLM5, IMG1, IMG2, MTF2, MRP10, MRPL28, RSM18, RPL29, RPL1B, RSM27, MRPL9, RRF1, MSR1, MRPL6, MEF2, RPL39, RSM26, MRP17, MRPL13, SLS1, TMA7, MRPL15, MRPL4, AEP1, MTG1, MRP7, MSK1, SWS2, MRPL10, IFM1, MSD1, MRP51, MRPL40, MSF1, MRPL51, MRP2 RNA metabolic process SLM5, MTF2, MBP1, MHR1, ARO80, RAI1, MSR1, NUC1, TOF2, DCS1, DSS1, HER2, MSK1, MRM1, VTS1, AEP3, SKS1, SGF11, RLM1, MSD1, MSF1 Transport MDM10, ERV46, YMC2, YSY6, MCH1, ATP17, AFG3, GLO3, FLC3, PCP1,YAP1802, OCT1, MMM1, ATP14, MSK1, MDM12, SKS1, ATP15 Generation of energy COX9, ATP17, QCR7, ABC1, COX6, PET10, SLS1, ATP14, COQ5, NDI1, PPA2, HER2, PCL8, ATP15, MRPL51 Lipid metabolic process CEM1, FAB1, ICT1, UPS1, LCB5, IZH4, SUR1, YDC1 Transcription MBP1, MHR1, ARO80, RAI1, TOF2, SKS1, SGF11, RLM1 DNA metabolic process RIM1, MBP1, RAD57, HFM1, NUC1, MGM101, IRC19, HMI1, MET7 Cellular amino acid metabolic process SLM5, ARO80, MSR1, HER2, MSK1, MSD1, ICL2, MSF1 Membrane organization MDM10, FZO1, GLO3, YAP1802, MMM1, MDM12 Ribosome biogenesis MRH4, RAI1, MRPL9, MTG1, MRM1 Response to stress TPS1, RAD57, MGM101, LCB5, YDC1 Cell cycle MBP1, RAD57, HFM1, SPO13, TOF2 Vesicle-mediated transport ERV46, GLO3, YAP1802 Carbohydrate metabolism TPS1, ANP1, CEM1, SKS1, SUR1, YPL088W, PCL8, ICL2 Signal transduction LCB5, RLM1 Protein modification process YSY6, COX9, ATP17, FPR2, QCR7, HSP31, ANP1, AFG3, PET117, FAB1, ABC1, PCP1, YAP1802, OCT1, VTS1, SKS1, MSF1, PBA1, OAZ1 Biological process unknown YBR144C, DEM1, YDL050C, PAR32, YDR157W, HRQ1, YDR509W, FPR2, HSP31, AIM10, CUE3, YGL218W, YGR219W, YGR283C, YJL119C, YJL132W, YJL152W, YJL193W, YKR012C, YMR187C, YMR310C, YNL011C, GEP3, CIR2, LCL1, YPL080C, YPR116W *The deletion strains were clustered based on their GO Process term using the GO Slim Mapper function at SGD. Mitochondria-associated genes are underlined and were identified from both the GO cellular component term clustering and the growth phenotype when plated on YPDG (0.2% dextrose, 3% glycerol). McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 8of9 Table S2. Rank list of deletion mutants resistant to Tcin* ORF Gene name 4 ␮M ratio 8 ␮M ratio 12 ␮M ratio 24 ␮M ratio Ratio sum BY4743 0.08 (0.15) 0.05 (0.09) 0.06 (0.32) 0.03 (0.31) 0.22 YHR014W SPO13 0.74 (0.37) 0.8 (0.01) 0.84 (0.11) 0.64 (0.15) 3.02 YLR193C UPS1 0.9 (0.09) 0.9 (0.07) 0.6 (0.05) 0.5 (0.78) 2.90 YLR262C-A TMA7 1.02 (0.07) 0.94 (0.05) 0.98 (0.01) 0.46 (0.47) 3.40 YKR010C TOF2 0.98 (0.14) 0.98 (0.04) 0.71 (0.09) 0.27 (0.76) 2.94 YLR099C ICT1 0.9 (0.21) 0.88 (0.01) 0.78 (0.09) 0.18 (0.7) 2.74 YNL011C YNL011C 0.99 (0.01) 0.93 (0.06) 0.62 (0.1) 0.13 (0.85) 2.67 YGR241C YAP1802 0.96 (0.12) 0.98 (0.12) 0.9 (0.08) 0.09 (1.54) 2.93 YDL054C MCH1 1.09 (0.06) 0.99 (0.03) 0.83 (0.03) 0.06 (0.74) 2.97 YOL101C IZH4 0.96 (0.02) 0.91 (0.03) 0.71 (0.45) 0.04 (0.37) 2.62 YMR187C YMR187C 0.84 (0.31) 0.94 (0.03) 0.73 (0.23) 0.02 (0.41) 2.53 YGL110C CUE3 0.92 (0.04) 0.85 (0.03) 0.71 (0.04) 0.02 (0.31) 2.50 YPL219W PCL8 0.89 (0.07) 0.87 (0.02) 0.56 (0.74) 0.01 (0.24) 2.33 YLR199C PBA1 0.94 (0.11) 0.91 (0.02) 0.63 (0.56) 0.01 (0.09) 2.49 YGL139W FLC3 0.98 (0.02) 0.84 (0.04) 0.62 (0.37) 0.01 (0.21) 2.46 YDL173W PAR32 0.92 (0.12) 0.94 (0.01) 0.59 (0.48) 0.01 (0.3) 2.45 *Liquid culture (OD600) growth ratios (toxin treatment divided by the control treatment) of the top 15 Tcin resistant deletion strains and the parental strain (BY4743) grown at four different concentrations of Tcin. The ratio sum score was calculated by summing the growth ratios across the different concentrations of Tcin. The ratio sum score provides an indication of the overall resistance of the strain and was used to rank resistance between strains. The coefficient of variation is presented in the parentheses. McLaughlin et al. www.pnas.org/cgi/content/short/0909777106 9of9.
Load more

Recommended publications
  • Supplementary Materials: Evaluation of Cytotoxicity and Α-Glucosidase Inhibitory Activity of Amide and Polyamino-Derivatives of Lupane Triterpenoids
    Supplementary Materials: Evaluation of cytotoxicity and α-glucosidase inhibitory activity of amide and polyamino-derivatives of lupane triterpenoids Oxana B. Kazakova1*, Gul'nara V. Giniyatullina1, Akhat G. Mustafin1, Denis A. Babkov2, Elena V. Sokolova2, Alexander A. Spasov2* 1Ufa Institute of Chemistry of the Ufa Federal Research Centre of the Russian Academy of Sciences, 71, pr. Oktyabrya, 450054 Ufa, Russian Federation 2Scientific Center for Innovative Drugs, Volgograd State Medical University, Novorossiyskaya st. 39, Volgograd 400087, Russian Federation Correspondence Prof. Dr. Oxana B. Kazakova Ufa Institute of Chemistry of the Ufa Federal Research Centre of the Russian Academy of Sciences 71 Prospeсt Oktyabrya Ufa, 450054 Russian Federation E-mail: [email protected] Prof. Dr. Alexander A. Spasov Scientific Center for Innovative Drugs of the Volgograd State Medical University 39 Novorossiyskaya st. Volgograd, 400087 Russian Federation E-mail: [email protected] Figure S1. 1H and 13C of compound 2. H NH N H O H O H 2 2 Figure S2. 1H and 13C of compound 4. NH2 O H O H CH3 O O H H3C O H 4 3 Figure S3. Anticancer screening data of compound 2 at single dose assay 4 Figure S4. Anticancer screening data of compound 7 at single dose assay 5 Figure S5. Anticancer screening data of compound 8 at single dose assay 6 Figure S6. Anticancer screening data of compound 9 at single dose assay 7 Figure S7. Anticancer screening data of compound 12 at single dose assay 8 Figure S8. Anticancer screening data of compound 13 at single dose assay 9 Figure S9. Anticancer screening data of compound 14 at single dose assay 10 Figure S10.
    [Show full text]
  • Role of Mitochondrial Ribosomal Protein S18-2 in Cancerogenesis and in Regulation of Stemness and Differentiation
    From THE DEPARTMENT OF MICROBIOLOGY TUMOR AND CELL BIOLOGY (MTC) Karolinska Institutet, Stockholm, Sweden ROLE OF MITOCHONDRIAL RIBOSOMAL PROTEIN S18-2 IN CANCEROGENESIS AND IN REGULATION OF STEMNESS AND DIFFERENTIATION Muhammad Mushtaq Stockholm 2017 All previously published papers were reproduced with permission from the publisher. Published by Karolinska Institutet. Printed by E-Print AB 2017 © Muhammad Mushtaq, 2017 ISBN 978-91-7676-697-2 Role of Mitochondrial Ribosomal Protein S18-2 in Cancerogenesis and in Regulation of Stemness and Differentiation THESIS FOR DOCTORAL DEGREE (Ph.D.) By Muhammad Mushtaq Principal Supervisor: Faculty Opponent: Associate Professor Elena Kashuba Professor Pramod Kumar Srivastava Karolinska Institutet University of Connecticut Department of Microbiology Tumor and Cell Center for Immunotherapy of Cancer and Biology (MTC) Infectious Diseases Co-supervisor(s): Examination Board: Professor Sonia Lain Professor Ola Söderberg Karolinska Institutet Uppsala University Department of Microbiology Tumor and Cell Department of Immunology, Genetics and Biology (MTC) Pathology (IGP) Professor George Klein Professor Boris Zhivotovsky Karolinska Institutet Karolinska Institutet Department of Microbiology Tumor and Cell Institute of Environmental Medicine (IMM) Biology (MTC) Professor Lars-Gunnar Larsson Karolinska Institutet Department of Microbiology Tumor and Cell Biology (MTC) Dedicated to my parents ABSTRACT Mitochondria carry their own ribosomes (mitoribosomes) for the translation of mRNA encoded by mitochondrial DNA. The architecture of mitoribosomes is mainly composed of mitochondrial ribosomal proteins (MRPs), which are encoded by nuclear genomic DNA. Emerging experimental evidences reveal that several MRPs are multifunctional and they exhibit important extra-mitochondrial functions, such as involvement in apoptosis, protein biosynthesis and signal transduction. Dysregulations of the MRPs are associated with severe pathological conditions, including cancer.
    [Show full text]
  • Supplementary Figures 1-14 and Supplementary References
    SUPPORTING INFORMATION Spatial Cross-Talk Between Oxidative Stress and DNA Replication in Human Fibroblasts Marko Radulovic,1,2 Noor O Baqader,1 Kai Stoeber,3† and Jasminka Godovac-Zimmermann1* 1Division of Medicine, University College London, Center for Nephrology, Royal Free Campus, Rowland Hill Street, London, NW3 2PF, UK. 2Insitute of Oncology and Radiology, Pasterova 14, 11000 Belgrade, Serbia 3Research Department of Pathology and UCL Cancer Institute, Rockefeller Building, University College London, University Street, London WC1E 6JJ, UK †Present Address: Shionogi Europe, 33 Kingsway, Holborn, London WC2B 6UF, UK TABLE OF CONTENTS 1. Supplementary Figures 1-14 and Supplementary References. Figure S-1. Network and joint spatial razor plot for 18 enzymes of glycolysis and the pentose phosphate shunt. Figure S-2. Correlation of SILAC ratios between OXS and OAC for proteins assigned to the SAME class. Figure S-3. Overlap matrix (r = 1) for groups of CORUM complexes containing 19 proteins of the 49-set. Figure S-4. Joint spatial razor plots for the Nop56p complex and FIB-associated complex involved in ribosome biogenesis. Figure S-5. Analysis of the response of emerin nuclear envelope complexes to OXS and OAC. Figure S-6. Joint spatial razor plots for the CCT protein folding complex, ATP synthase and V-Type ATPase. Figure S-7. Joint spatial razor plots showing changes in subcellular abundance and compartmental distribution for proteins annotated by GO to nucleocytoplasmic transport (GO:0006913). Figure S-8. Joint spatial razor plots showing changes in subcellular abundance and compartmental distribution for proteins annotated to endocytosis (GO:0006897). Figure S-9. Joint spatial razor plots for 401-set proteins annotated by GO to small GTPase mediated signal transduction (GO:0007264) and/or GTPase activity (GO:0003924).
    [Show full text]
  • 1 AGING Supplementary Table 2
    SUPPLEMENTARY TABLES Supplementary Table 1. Details of the eight domain chains of KIAA0101. Serial IDENTITY MAX IN COMP- INTERFACE ID POSITION RESOLUTION EXPERIMENT TYPE number START STOP SCORE IDENTITY LEX WITH CAVITY A 4D2G_D 52 - 69 52 69 100 100 2.65 Å PCNA X-RAY DIFFRACTION √ B 4D2G_E 52 - 69 52 69 100 100 2.65 Å PCNA X-RAY DIFFRACTION √ C 6EHT_D 52 - 71 52 71 100 100 3.2Å PCNA X-RAY DIFFRACTION √ D 6EHT_E 52 - 71 52 71 100 100 3.2Å PCNA X-RAY DIFFRACTION √ E 6GWS_D 41-72 41 72 100 100 3.2Å PCNA X-RAY DIFFRACTION √ F 6GWS_E 41-72 41 72 100 100 2.9Å PCNA X-RAY DIFFRACTION √ G 6GWS_F 41-72 41 72 100 100 2.9Å PCNA X-RAY DIFFRACTION √ H 6IIW_B 2-11 2 11 100 100 1.699Å UHRF1 X-RAY DIFFRACTION √ www.aging-us.com 1 AGING Supplementary Table 2. Significantly enriched gene ontology (GO) annotations (cellular components) of KIAA0101 in lung adenocarcinoma (LinkedOmics). Leading Description FDR Leading Edge Gene EdgeNum RAD51, SPC25, CCNB1, BIRC5, NCAPG, ZWINT, MAD2L1, SKA3, NUF2, BUB1B, CENPA, SKA1, AURKB, NEK2, CENPW, HJURP, NDC80, CDCA5, NCAPH, BUB1, ZWILCH, CENPK, KIF2C, AURKA, CENPN, TOP2A, CENPM, PLK1, ERCC6L, CDT1, CHEK1, SPAG5, CENPH, condensed 66 0 SPC24, NUP37, BLM, CENPE, BUB3, CDK2, FANCD2, CENPO, CENPF, BRCA1, DSN1, chromosome MKI67, NCAPG2, H2AFX, HMGB2, SUV39H1, CBX3, TUBG1, KNTC1, PPP1CC, SMC2, BANF1, NCAPD2, SKA2, NUP107, BRCA2, NUP85, ITGB3BP, SYCE2, TOPBP1, DMC1, SMC4, INCENP. RAD51, OIP5, CDK1, SPC25, CCNB1, BIRC5, NCAPG, ZWINT, MAD2L1, SKA3, NUF2, BUB1B, CENPA, SKA1, AURKB, NEK2, ESCO2, CENPW, HJURP, TTK, NDC80, CDCA5, BUB1, ZWILCH, CENPK, KIF2C, AURKA, DSCC1, CENPN, CDCA8, CENPM, PLK1, MCM6, ERCC6L, CDT1, HELLS, CHEK1, SPAG5, CENPH, PCNA, SPC24, CENPI, NUP37, FEN1, chromosomal 94 0 CENPL, BLM, KIF18A, CENPE, MCM4, BUB3, SUV39H2, MCM2, CDK2, PIF1, DNA2, region CENPO, CENPF, CHEK2, DSN1, H2AFX, MCM7, SUV39H1, MTBP, CBX3, RECQL4, KNTC1, PPP1CC, CENPP, CENPQ, PTGES3, NCAPD2, DYNLL1, SKA2, HAT1, NUP107, MCM5, MCM3, MSH2, BRCA2, NUP85, SSB, ITGB3BP, DMC1, INCENP, THOC3, XPO1, APEX1, XRCC5, KIF22, DCLRE1A, SEH1L, XRCC3, NSMCE2, RAD21.
    [Show full text]
  • Identification of Gold Nanoparticle-Resistant Mutants of Saccharomyces Cerevisiae Suggests a Role for Respiratory Metabolism in Mediating Toxicity
    Identification of Gold Nanoparticle-Resistant Mutants of Saccharomyces cerevisiae Suggests a Role for Respiratory Metabolism in Mediating Toxicity Smith, M. R., Boenzli, M. G., Hindagolla, V., Ding, J., Miller, J. M., Hutchison, J. E., . Bakalinsky, A. T. (2013). Identification of gold nanoparticle-resistant mutants of saccharomyces cerevisiae suggests a role for respiratory metabolism in mediating toxicity. Applied and Environmental Microbiology, 79(2), 728-733. doi: 10.1128/AEM.01737-12 10.1128/AEM.01737-12 American Society for Microbiology Version of Record http://cdss.library.oregonstate.edu/sa-termsofuse Identification of Gold Nanoparticle-Resistant Mutants of Saccharomyces cerevisiae Suggests a Role for Respiratory Metabolism in Mediating Toxicity Mark R. Smith,a Matthew G. Boenzli,a Vihangi Hindagolla,a Jun Ding,a,b John M. Miller,c,d James E. Hutchison,c,d Jeffrey A. Greenwood,b Hagai Abeliovich,e Alan T. Bakalinskya Downloaded from Department of Food Science and Technology, Oregon State University, Corvallis, Oregon, USAa; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon, USAb; The Safer Nanomaterials and Nanomanufacturing Initiative, Eugene, Oregon, USAc; Department of Chemistry and the Materials Science Institute, University of Oregon, Eugene, Oregon, USAd; Department of Biochemistry and Food Science, Hebrew University of Jerusalem, Rehovot, Israele Positively charged gold nanoparticles (0.8-nm core diameter) reduced yeast survival, but not growth, at a concentration of 10 to 100 ␮g/ml. Among 17 resistant deletion mutants isolated in a genome-wide screen, highly significant enrichment was observed for respiration-deficient mutants lacking genes encoding proteins associated with the mitochondrion. http://aem.asm.org/ he increasing use of nanomaterials in industrial processes and Open Biosystems, Inc.) was screened for resistance to these gold Tcommercial products has generated a need for systematic as- nanoparticles.
    [Show full text]
  • Stem Cells® Original Article
    ® Stem Cells Original Article Properties of Pluripotent Human Embryonic Stem Cells BG01 and BG02 XIANMIN ZENG,a TAKUMI MIURA,b YONGQUAN LUO,b BHASKAR BHATTACHARYA,c BRIAN CONDIE,d JIA CHEN,a IRENE GINIS,b IAN LYONS,d JOSEF MEJIDO,c RAJ K. PURI,c MAHENDRA S. RAO,b WILLIAM J. FREEDa aCellular Neurobiology Research Branch, National Institute on Drug Abuse, Department of Health and Human Services (DHHS), Baltimore, Maryland, USA; bLaboratory of Neuroscience, National Institute of Aging, DHHS, Baltimore, Maryland, USA; cLaboratory of Molecular Tumor Biology, Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA; dBresaGen Inc., Athens, Georgia, USA Key Words. Embryonic stem cells · Differentiation · Microarray ABSTRACT Human ES (hES) cell lines have only recently been compared with pooled human RNA. Ninety-two of these generated, and differences between human and mouse genes were also highly expressed in four other hES lines ES cells have been identified. In this manuscript we (TE05, GE01, GE09, and pooled samples derived from describe the properties of two human ES cell lines, GE01, GE09, and GE07). Included in the list are genes BG01 and BG02. By immunocytochemistry and reverse involved in cell signaling and development, metabolism, transcription polymerase chain reaction, undifferenti- transcription regulation, and many hypothetical pro- ated cells expressed markers that are characteristic of teins. Two focused arrays designed to examine tran- ES cells, including SSEA-3, SSEA-4, TRA-1-60, TRA-1- scripts associated with stem cells and with the 81, and OCT-3/4. Both cell lines were readily main- transforming growth factor-β superfamily were tained in an undifferentiated state and could employed to examine differentially expressed genes.
    [Show full text]
  • Cardiac SARS‐Cov‐2 Infection Is Associated with Distinct Tran‐ Scriptomic Changes Within the Heart
    Cardiac SARS‐CoV‐2 infection is associated with distinct tran‐ scriptomic changes within the heart Diana Lindner, PhD*1,2, Hanna Bräuninger, MS*1,2, Bastian Stoffers, MS1,2, Antonia Fitzek, MD3, Kira Meißner3, Ganna Aleshcheva, PhD4, Michaela Schweizer, PhD5, Jessica Weimann, MS1, Björn Rotter, PhD9, Svenja Warnke, BSc1, Carolin Edler, MD3, Fabian Braun, MD8, Kevin Roedl, MD10, Katharina Scher‐ schel, PhD1,12,13, Felicitas Escher, MD4,6,7, Stefan Kluge, MD10, Tobias B. Huber, MD8, Benjamin Ondruschka, MD3, Heinz‐Peter‐Schultheiss, MD4, Paulus Kirchhof, MD1,2,11, Stefan Blankenberg, MD1,2, Klaus Püschel, MD3, Dirk Westermann, MD1,2 1 Department of Cardiology, University Heart and Vascular Center Hamburg, Germany. 2 DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck. 3 Institute of Legal Medicine, University Medical Center Hamburg‐Eppendorf, Germany. 4 Institute for Cardiac Diagnostics and Therapy, Berlin, Germany. 5 Department of Electron Microscopy, Center for Molecular Neurobiology, University Medical Center Hamburg‐Eppendorf, Germany. 6 Department of Cardiology, Charité‐Universitaetsmedizin, Berlin, Germany. 7 DZHK (German Centre for Cardiovascular Research), partner site Berlin, Germany. 8 III. Department of Medicine, University Medical Center Hamburg‐Eppendorf, Germany. 9 GenXPro GmbH, Frankfurter Innovationszentrum, Biotechnologie (FIZ), Frankfurt am Main, Germany. 10 Department of Intensive Care Medicine, University Medical Center Hamburg‐Eppendorf, Germany. 11 Institute of Cardiovascular Sciences,
    [Show full text]
  • The C-Terminal Extension Domain of Saccharomyces Cerevisiae Mrpl32, a Homolog of Ribosomal Protein L32, Functions in Trans to Support Mitochondrial Translation
    Genes Genet. Syst. (2018) 93, p. 21–24 Function of C-terminal extension of MrpL32 21 The C-terminal extension domain of Saccharomyces cerevisiae MrpL32, a homolog of ribosomal protein L32, functions in trans to support mitochondrial translation Ivo Ngundu Woogeng and Madoka Kitakawa* Department of Biology, Faculty of Science, Kobe University, Rokko 1-1, Kobe, Hyogo 657-8501, Japan (Received 3 June 2017, accepted 10 August 2017; J-STAGE Advance published date: 17 January 2018) Mitochondrial ribosomal protein L32 (MrpL32) of Saccharomyces cerevisiae is homologous to the bacterial L32 ribosomal protein. MrpL32 carries an N-terminal mitochondrion-targeting sequence (MTS) and is about 60 amino acid residues lon- ger at the C-terminus. Adding to its function as a leader sequence, the MTS of MrpL32 has been reported to regulate ribosome biogenesis through its processing by m-AAA protease. However, the function of the C-terminal extension (CE) remains totally unknown. Therefore, we constructed a series of C-terminally truncated mrpl32 (mrpl32ΔC) genes and expressed them in a Δmrpl32 mutant to examine their function. Interestingly, some MrpL32ΔC derivatives exhibited temperature- sensitive (ts) growth on medium with non-fermentable carbon sources. Further- more, the CE domain of MrpL32, expressed separately from MrpL32ΔC, could rescue the ts phenotype of mutants by improving mitochondrial protein synthesis. Key words: C-terminal extension, mitochondrial ribosome, MrpL32 Many protein components of mitochondrial ribosomes (Huff et al., 1993). are homologous to eubacterial ribosomal proteins, but they MrpL32 is unique; its long mitochondrion-targeting tend to be larger than the corresponding bacterial proteins sequence (MTS) (71 aa) is a target of m-AAA prote- (Graack and Wittmann-Liebold, 1998; Gan et al., 2002; ase, and the loss of MrpL32 processing is thoroughly Smits et al., 2007).
    [Show full text]
  • Supplementary Dataset S2
    mitochondrial translational termination MRPL28 MRPS26 6 MRPS21 PTCD3 MTRF1L 4 MRPL50 MRPS18A MRPS17 2 MRPL20 MRPL52 0 MRPL17 MRPS33 MRPS15 −2 MRPL45 MRPL30 MRPS27 AURKAIP1 MRPL18 MRPL3 MRPS6 MRPS18B MRPL41 MRPS2 MRPL34 GADD45GIP1 ERAL1 MRPL37 MRPS10 MRPL42 MRPL19 MRPS35 MRPL9 MRPL24 MRPS5 MRPL44 MRPS23 MRPS25 ITB ITB ITB ITB ICa ICr ITL original ICr ICa ITL ICa ITL original ICr ITL ICr ICa mitochondrial translational elongation MRPL28 MRPS26 6 MRPS21 PTCD3 MRPS18A 4 MRPS17 MRPL20 2 MRPS15 MRPL45 MRPL52 0 MRPS33 MRPL30 −2 MRPS27 AURKAIP1 MRPS10 MRPL42 MRPL19 MRPL18 MRPL3 MRPS6 MRPL24 MRPS35 MRPL9 MRPS18B MRPL41 MRPS2 MRPL34 MRPS5 MRPL44 MRPS23 MRPS25 MRPL50 MRPL17 GADD45GIP1 ERAL1 MRPL37 ITB ITB ITB ITB ICa ICr original ICr ITL ICa ITL ICa ITL original ICr ITL ICr ICa translational termination MRPL28 MRPS26 6 MRPS21 PTCD3 C12orf65 4 MTRF1L MRPL50 MRPS18A 2 MRPS17 MRPL20 0 MRPL52 MRPL17 MRPS33 −2 MRPS15 MRPL45 MRPL30 MRPS27 AURKAIP1 MRPL18 MRPL3 MRPS6 MRPS18B MRPL41 MRPS2 MRPL34 GADD45GIP1 ERAL1 MRPL37 MRPS10 MRPL42 MRPL19 MRPS35 MRPL9 MRPL24 MRPS5 MRPL44 MRPS23 MRPS25 ITB ITB ITB ITB ICa ICr original ICr ITL ICa ITL ICa ITL original ICr ITL ICr ICa translational elongation DIO2 MRPS18B MRPL41 6 MRPS2 MRPL34 GADD45GIP1 4 ERAL1 MRPL37 2 MRPS10 MRPL42 MRPL19 0 MRPL30 MRPS27 AURKAIP1 −2 MRPL18 MRPL3 MRPS6 MRPS35 MRPL9 EEF2K MRPL50 MRPS5 MRPL44 MRPS23 MRPS25 MRPL24 MRPS33 MRPL52 EIF5A2 MRPL17 SECISBP2 MRPS15 MRPL45 MRPS18A MRPS17 MRPL20 MRPL28 MRPS26 MRPS21 PTCD3 ITB ITB ITB ITB ICa ICr ICr ITL original ITL ICa ICa ITL ICr ICr ICa original
    [Show full text]
  • A Homozygous MRPL24 Mutation Causes a Complex Movement Disorder and Affects the Mitoribosome Assembly T
    Neurobiology of Disease 141 (2020) 104880 Contents lists available at ScienceDirect Neurobiology of Disease journal homepage: www.elsevier.com/locate/ynbdi A homozygous MRPL24 mutation causes a complex movement disorder and affects the mitoribosome assembly T Michela Di Nottiaa,1, Maria Marcheseb,1, Daniela Verrignia, Christian Daniel Muttic, Alessandra Torracoa, Romina Olivad, Erika Fernandez-Vizarrac, Federica Moranib, Giulia Trania, Teresa Rizzaa, Daniele Ghezzie,f, Anna Ardissoneg,h, Claudia Nestib, Gessica Vascoi, Massimo Zevianic, Michal Minczukc, Enrico Bertinia, Filippo Maria Santorellib,2, ⁎ Rosalba Carrozzoa, ,2 a Unit of Muscular and Neurodegenerative Disorders, Laboratory of Molecular Medicine, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy b Molecular Medicine & Neurogenetics, IRCCS Fondazione Stella Maris, Pisa, Italy c MRC Mitochondrial Biology Unit, University of Cambridge, Cambridge, UK d Department of Sciences and Technologies, University Parthenope of Naples, Naples, Italy e Unit of Medical Genetics and Neurogenetics, Fondazione IRCCS Istituto Neurologico Carlo Besta, Milan, Italy f Department of Pathophysiology and Transplantation, University of Milan, Milan, Italy g Child Neurology Unit, Fondazione IRCCS Istituto Neurologico Carlo Besta, Milan, Italy h Department of Molecular and Translational Medicine DIMET, University of Milan-Bicocca, Milan, Italy i Department of Neurosciences, IRCCS Bambino Gesù Children Hospital, Rome, Italy ARTICLE INFO ABSTRACT Keywords: Mitochondrial ribosomal protein large 24 (MRPL24) is 1 of the 82 protein components of mitochondrial ribo- Mitochondrial disorders somes, playing an essential role in the mitochondrial translation process. Movement disorder We report here on a baby girl with cerebellar atrophy, choreoathetosis of limbs and face, intellectual dis- MRPL24 ability and a combined defect of complexes I and IV in muscle biopsy, caused by a homozygous missense mu- Mitoribosomes tation identified in MRPL24.
    [Show full text]
  • Influenza-Specific Effector Memory B Cells Predict Long-Lived Antibody Responses to Vaccination in Humans
    bioRxiv preprint doi: https://doi.org/10.1101/643973; this version posted February 18, 2021. 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. Influenza-specific effector memory B cells predict long-lived antibody responses to vaccination in humans Anoma Nellore1, Esther Zumaquero2, Christopher D. Scharer3, Rodney G. King2, Christopher M. Tipton4, Christopher F. Fucile5, Tian Mi3, Betty Mousseau2, John E. Bradley6, Fen Zhou2, Paul A. Goepfert1, Jeremy M. Boss3, Troy D. Randall6, Ignacio Sanz4, Alexander F. Rosenberg2,5, Frances E. Lund2 1Dept. of Medicine, Division of Infectious Disease, 2Dept of Microbiology, 5Informatics Institute, 6Dept. of Medicine, Division of Clinical Immunology and Rheumatology and at The University of Alabama at Birmingham, Birmingham, AL 35294 USA 3Dept. of Microbiology and Immunology and 4Department of Medicine, Division of Rheumatology Emory University, Atlanta, GA 30322, USA Correspondence should be addressed to: Frances E. Lund, PhD Charles H. McCauley Professor and Chair Dept of Microbiology University of Alabama at Birmingham 276 BBRB Box 11 1720 2nd Avenue South Birmingham AL 35294-2170 [email protected] SHORT RUNNING TITLE: Effector memory B cell development after influenza vaccination 1 bioRxiv preprint doi: https://doi.org/10.1101/643973; this version posted February 18, 2021. 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. Abstract Seasonal influenza vaccination elicits hemagglutinin (HA)-specific CD27+ memory B cells (Bmem) that differ in expression of T-bet, BACH2 and TCF7.
    [Show full text]
  • AFG3L2 Supports Mitochondrial Protein Synthesis and Purkinje Cell Survival
    AFG3L2 supports mitochondrial protein synthesis and Purkinje cell survival Eva R. Almajan, … , Thomas Langer, Elena I. Rugarli J Clin Invest. 2012;122(11):4048-4058. https://doi.org/10.1172/JCI64604. Research Article Neuroscience Mutations in the AFG3L2 gene have been linked to spinocerebellar ataxia type 28 and spastic ataxia-neuropathy syndrome in humans; however, the pathogenic mechanism is still unclear. AFG3L2 encodes a subunit of the mitochondrial m-AAA protease, previously implicated in quality control of misfolded inner mitochondrial membrane proteins and in regulatory functions via processing of specific substrates. Here, we used a conditional Afg3l2 mouse model that allows restricted deletion of the gene in Purkinje cells (PCs) to shed light on the pathogenic cascade in the neurons mainly affected in the human diseases. We demonstrate a cell-autonomous requirement of AFG3L2 for survival of PCs. Examination of PCs prior to neurodegeneration revealed fragmentation and altered distribution of mitochondria in the dendritic tree, indicating that abnormal mitochondrial dynamics is an early event in the pathogenic process. Moreover, PCs displayed features pointing to defects in mitochondrially encoded respiratory chain subunits at early stages. To unravel the underlying mechanism, we examined a constitutive knockout of Afg3l2, which revealed a decreased rate of mitochondrial protein synthesis associated with impaired mitochondrial ribosome assembly. We therefore propose that defective mitochondrial protein synthesis, leading to early-onset fragmentation of the mitochondrial network, is a central causative factor in AFG3L2-related neurodegeneration. Find the latest version: https://jci.me/64604/pdf Research article AFG3L2 supports mitochondrial protein synthesis and Purkinje cell survival Eva R. Almajan,1 Ricarda Richter,2 Lars Paeger,1 Paola Martinelli,1 Esther Barth,1 Thorsten Decker,2 Nils-Göran Larsson,3,4 Peter Kloppenburg,1,4,5 Thomas Langer,2,3,4,5 and Elena I.
    [Show full text]