Vol. 6 No. 2 June 2001
| Volume 6 (2001) pp 117-140 | |
| Title | POLY(ADP-RIBOSE) POLYMERASE-1 REGULATES THE STABILITY OF THE WILD-TYPE p53 PROTEIN |
| Authors | Jozefa Wesierska-Gadek* and Gerald Schmid# |
| Abstract | We investigated the interaction between poly(ADP-ribose) polymerase-1 (PARP-1) and the product of the tumor suppressor gene p53 using two different approaches. In the first approach, we used primary and immortalized cells derived from wt and PARP-1 -/- mice. We examined whether PARP-1 deficiency would affect the expression of the wild-type (wt) p53 protein. The inactivation of the PARP-1 gene markedly affected the constitutive expression of the wt p53 protein. Interestingly, only the regularly spliced form of wt p53 was reduced to a barely detectable level in consequence to an approximately 8-fold shortening of its half-life, whereas the level of alternatively spliced p53 remained unchanged. Moreover, reconstitution of cells lacking the PARP-1 gene with the human counterpart restored the normal stability of the regularly spliced p53 protein. In the second approach, we performed experiments with c-Ha-ras transformed primary rat cells overexpressing the p53135val. mutant alone or in combination with PARP-1. The advantage of this temperature sensitive p53135val mutant is its oncogenic character at 37°C, connected with cytoplasmic localization of p53, and its tumor suppressor activity at 32°C, accompanied by p53 translocation into the nucleus. No noticeable differences in proliferation and G1 accumulationwere observed between cells expressing p53135val with or without PARP-1. On the other hand, a comparison of the recovery of G1 arrested cells after a shift up to 37°C for both cell lines showed dramatic differences in the kinetics. While cells expressing p53135val rapidly reached the characteristic S-phase level after a shift up to basal temperature, cells additionally expressing PARP-1 rested in G1 despite the temperature elevation. This coincided with exclusively cytoplasmic p53 protein in cells expressing p53135val and predominantly nuclear localization of p53 in p53135val +PARP-1 cells, as evidenced by immunostaining. Determination of the p53 level during the maintenance of cells at 32°C revealed a marked decrease in the level of p53 in cells expressing p53135val alone, whereas in cells coexpressing PARP-1, the level of p53 remained largely unaffected. This indicates that the stability of wild-type p53 greatly differed between both cell lines. Furthermore, the inhibition of PARP-1 activity in G1 arrested cells by 3-aminobenzamide abolished its stabilizing effect on the wildtype p53 protein. Taken together, our results indicate that PARP-1 regulates the stability of the wt p53 protein and that its enzymatic activity is necessary for this stabilizing action. |
| Address and Contact Information | Institute of Cancer Research, University of Vienna, Borschkegasse 8 a,
A-1090 Vienna, Austria *Corresponding author: Jozefa Wesierska-Gadek, Phone 43-1-4277 ext. 65247 Fax 43-1-4277 ext. 65194 or 965 1 e-mail: Jozefa.Antonia.Gadek-Wesierski@univie.ac.at #present address: International Agency for Research on Cancer |
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| Volume 6 (2001) pp 141-159 | |
| Title | A COMPARISON OF THE METHODS APPLIED TO DETECT APOPTOSIS IN GENOTOXICALLY-DAMAGED LYMPHOCYTES CULTURED IN THE PRESENCE OF FOUR ANTIMUTAGENS |
| Authors | Kazimierz Gasiorowski1*, Barbara Brokos1, Anna Kulma2, Antoni Ogorzalek3 and Katarzyna Skorkowska1 |
| Abstract | The sensitivity of the available methods of apoptosis detection in lymphocyte cultures was tested. Cells were preincubated with genotoxic agents: hydrogen peroxide (0.2 mM; 20min.) and benzo[a]pyrene (40 µM;90min.), and then cultured for 36h in the presence of a lectin (PHA-M;1%v/v) and one of the following potentially antimutagenic agents: alkylresorcinols, anthocyanins, todralazine and fluphenazine. It was established that staining with a mixture of fluorochromes (ethidium bromide and acridine orange) provided the highest amount of detected apoptotic cells, and the best repeatability of the results in subsequent experiments. Calculation of the Spearman's rank correlation coefficients proved that there was a high correlation between the results obtained by the ethidium bromide/acridine orange method and those obtained by identifying genomic DNA fragmentation by means of FIGE-electrophoresis. Therefore, these two methods were chosen for further studies of the tested antimutagens’ impact on apoptosis in genotoxically-damaged lymphocytes. |
| Address and Contact Information | 1Wroclaw Medical University, Department of Basic Medical Sciences,
Kochanowskiego 14, 51- 601 Wroclaw, Poland, 2University of Wroclaw, Department of Genetic Biochemistry, Przybyszewskiego 63/77, 51-148 Wroclaw, Poland, 3University of Wroclaw, Department of Zoology, ul.Sienkiewicza 21, 50-335 Wroclaw, Poland *Corresponding author, fax: (+48 71) 3479211, e-mail: kaz@basmed.am.wroc.pl |
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| Volume 6 (2001) pp 161-165 | |
| Title | OXYETHYLENE CHAIN-CATION COMPLEXATION; NONIONIC POLYOXYETHYLENE DETERGENTS ATTAIN A POSITIVE CHARGE AND DEMONSTRATE ELECTROSTATIC HEAD GROUP INTERACTIONS |
| Authors | Henry Hägerstrand1, Johan Bobacka2, Malgorzata Bobrowska-Hägerstrand1, Veronika Kralj-Iglic3, Miha Fošnaric4 and Aleš Iglic4 |
| Abstract | We report literature data indicating that the polyoxyethylene chain of polyoxyethylene detergents attracts cations via dipole-ion interactions thereby attaining a positive charge character. This implies that nonionic polyoxyethylene detergents like Triton X-100 and C12E8 may interact electrostatically with phospholipid head groups. We describe how a positive charge character of Triton X-100 and C12E8 can explain their hitherto mysterious stomatocytogenic shape altering effect in human erythrocytes. |
| Address and Contact Information | 1Department of Biology and 2Laboratory of Analytical Chemistry, Abo Akademi
University, FIN-20520, Abo/Turku, Finland, 3Institute of Biophysics, Medical Faculty, University of Ljubljana and Clinical Centre Ljubljana and 4Laboratory of Applied Physics, Faculty of Electrical Engineering, University of Ljubljana, SI- 1000, Ljubljana, Slovenia |
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| Volume 6 (2001) pp 167-171 | |
| Title | ON STABILITY OF CIRCULAR HOLE IN MEMBRANE BILAYER |
| Authors | Miha Fo©naric1, Veronika Kralj-Iglic2, Henry HÄgerstrand3 and Ale© Iglic1 |
| Abstract | It was observed recently that nonionic surfactant octaethyleneglycol dodecylether (C12E8) decreases threshold for irreversible electroporation in membrane bilayers. In accordance, it is shown theoretically in this work that anisotropic C12E8 membrane inclusions may stabilize circular hole in a flat membrane segment. |
| Address and Contact Information | 1Laboratory of Applied Physics, Faculty of Electrical Engineering, University
of Ljubljana, SI-1000, Ljubljana, Slovenia, 2Institute of Biophysics, Faculty of Medicine, University of Ljubljana, SI-1000, Ljubljana, Slovenia, 3Department of Biology, Abo Akademi University, FIN-20520, Abo/Turku, Finland |
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| Volume 6 (2001) pp 173-184 | |
| Title | INHIBITION OF p38 KINASE MIMICS SURVIVAL SIGNAL-LINKED PROTECTION AGAINST APOPTOSIS IN RAT CEREBELLAR GRANULE NEURONS |
| Authors | Rathna Nath1*, Kim Mcginnis3, Satavisha Dutta1,2, Brenda Shivers2 And Kevin K.W. Wang1 |
| Abstract | The mitogen-activated protein kinase (MAPK) cascades are thought to be important mediators in the transduction of extracellular signals into cellular responses. The p38 kinase, a member of the MAPK superfamily, is activated by a wide variety of extracellular stimuli and has been implicated in neuronal apoptosis induced by glutamate. In this study we have examined the role of p38 kinase in the potassium deprivation model of apoptosis in rat cerebellar granule neurons (CGN). An increase in p38 kinase activity was observed with a 15-minute potassium deprivation when compared to the basal level. We also found that SB203580 and PD169316, specific p38 kinase inhibitors, significantly attenuated apoptosis in potassium-deprived cells in a dose dependent manner. A decrease in caspase-3 mediated DEVD-MCA , substrate hydrolysis and the appearance of the 120 kDa-spectrin breakdown product in cells treated with SB203580 further suggests that the p38 kinase acts upstream of caspase-3 in the apoptosis cascade. The data provides evidence for an essential role of p38 kinase in mediating apoptotic cell death in CGN and the inhibition of p38 kinase mimics the suppression of apoptosis provided by natural survival signals. |
| Address and Contact Information | Depts. of Neuroscience Therapeutics1 and Cell Biology 2, Pfizer Global Research and Development, 2800 Plymouth Road, Ann Arbor, MI 48105; 3Current address: Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129, USA |
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| Volume 5 (2000) pp 185-234 | |
| Seminar Title | W. MEJBAUM-KATZENELLENBOGEN'S MOLECULAR BIOLOGY SEMINARS: 8. MEMBRANE SKELETON AND ITS REGULATORY FUNCTIONS |
| Abstracts List |
ROLE OF GOLGI MATRIX PROTEINS IN REGULATING SECRETORY TRAFFIC Cecilia Alvarez and Elizabeth Sztul - p.186 STABILITY OF THE SPECTRIN REPEAT UNITS Lars Backman and Lars Löfvenberg - p.187 ISOFORMS, DISTRIBUTION AND ACTIVITIES OF ? SPECTRINS AND 4.1 PROTEINS IN BRAIN Anthony J. Baines, Paola A. Bignone, Nandini V. L. Hayes, Lisa A. Keating, Gareth W. Phillips and Catherine Scott - p.188 ON THE ROLE OF SPECTRIN AND SPECTRIN SH3 DOMAIN IN INTEGRIN -INDUCED SIGNALING Katarzyna Białkowska and Joan E. B. Fox - p.189 ASSOCIATION OF A SPECTRIN-LIKE PROTEIN WITH AN APICAL ACTIN-ORGANIZED ENDOPLASMIC-RETICULUM AGGREGATE AND ITS PUTATIVE FUNCTIONS IN GRAVITROPICALLY TIP-GROWING PLANT CELLS Markus Braun - p.190 TALIN DISTRIBUTION DURING DIFFERENTIATION OF SATELLITE CELLS ISOLATED FROM M. EDL AND M. SOLEUS TREATED BY PHORBOL ESTER Edyta Brzoska, Edyta Wróbel and Jerzy Moraczewski - p.191 DOLICHOLS AND POLYPRENOLS; ELEMENTS OF MEMBRANES, COENZYMES AND SECONDARY METABOLITES Tadeusz Chojnacki, Wiesław Jankowski and Ewa Świeżewska - p.192 ELECTROPHORETIC ANALYSIS OF ALBUMINS AND GLOBULINS ISOLATED FROM THE WINTER TRITICALE OF DIFFERENT RESISTANCE TO THE GRAIN APHID Antoni Piotr Ciepiela and Iwona Sprawka - p.193 NUCLEAR AND CYTOPLASMIC LOCALIZATION SIGNALS IN PROTEIN 4.1R Isabel Correas, Carmen M. Pérez-Ferreiro, María-José Lallena and Carlos M. Luque - p.195 MECHANICS OF STACKED MEMBRANE COMPARTMENTS Jure Derganc, Bojan Božič, Boštjan Žekš and Saša Svetina - p.196 APICAL SPECTRIN LIKE PROTEIN AND SUBAPICAL FINE BUNDLES OF ACTIN FILAMENTS CORRELATE WITH PLANT CELL TIP GROWTH Norbert De Ruijter, Tijs Ketelaar and Anne Mie Emons - p.197 INTERACTION OF BRAIN AND ERYTHROCYTE SPECTRIN WITH ANIONIC PHOSPHOLIPIDS Witold Diakowski and Aleksander F. Sikorski - p.198
STRUCTURAL STUDIES ON CYTOSKELETAL PROTEINS Kristina Djinovič-Carugo - p.199 THE EFFECT OF CHEMOTHERAPY WITH FLUODARABINE/MITO-XANTRONE/DEXAMETHASONE ON THE DISTRIBUTION OF SPECTRIN IN LYMPHOCYTES OF NON-HODGKIN LYMPHOMA PATIENTS Patrycja Dubielecka, Stanisław Potoczek, Bożena Jaźwiec, Joanna Miłoszewska, Kazimierz Kuliczkowski and Aleksander F. Sikorski - p.200 STRAIN FIELD MAPPING OF THE CELL MEMBRANE OF ENDOTHELIAL CELLS Wolfgang Feneberg and Erich Sackmann - p.201 ANKYRINS AND ORGANIZATION OF MEMBRANE DOMAINS IN MUSCLE Claire Gagelin, Bruno Constantin, Christiane Deprette, Marie-Aline Ludosky and Ekaterini Kordeli - p.202 IDENTIFICATION AND LOCALISATION OF CALDESMON-LIKE PROTEIN IN AMOEBA PROTEUS Małgorzata Gągola, Robert Makuch, Anna Łopatowska and Wanda Kłopocka - p.203 ANNEXINS AND MEMBRANE ORGANISATION IN THE ENDOCYTIC PATHWAY Völker Gerke - p.204 ERYTHROCYTE SPECTRIN IS AN E2 UBIQUITIN CONJUGATING ENZYME Steven R. Goodman, David G. Kakhniashvili, JosĂ©, Sangerman, Tanuja Chaudhary, Warren E. Zimmer, F. Aladar Bencsath and Ian Jardine - p.205 AMPHIPHILE-INDUCED VESICULATION IN AGED HEREDITARY SPHERO-CYTOSIS ERYTHROCYTES INDICATES NORMAL MEMBRANE STABILITY PROPERTIES UNDER NON-STARVING CONDITIONS Henry Hägerstrand, Aleš Iglič and Stefan Eber - p.206 SPECTRIN MOTIFS ARE DETECTED IN PLANT AND YEAST GENOMES Beata Hanus-Lorenz, Anita Hryniewicz-Jankowska, Jacek Leluk, Michał Lorenz, Jacek Skała and Aleksander F. Sikorski - p.207 THE DYNAMIC CYTOSKELETON OF THE MEGAKARYTOCYTE John H. Hartwig, Kurt Barkalow and Joe Italiano, Jr - p.208 THE TWO MAJOR PROTEIN ISOFORMS OF ANKYRIN 2 ARE DIFFEREN-TIALLY LOCALIZED IN DROSOPHILA NEURONS Michael Hortsch, Kerry L. Paisley, Ming-Zhu Tian,Min Qian, Michael Bouley and Robert Chandler - p.209 CHANGES OF MEMBRANE SKELETON OF DENDRITIC CELLS DURING DIFFERENTIATION Jerzy Kawiak - p.210 EFFECTS OF BLOCKING OF ENDOGENOUS RHO FAMILY GTP-BINDING PROTEINS ON MORPHOLOGY, ADHESION AND LOCOMOTION OF AMOEBA PROTEUS Wanda Kłopocka and M. Jolanta Rędowicz - p.211 ERYTHROCYTE RESPONSE TO NEAR INFRARED RADIATION Małgorzata Komorowska, Arnaud Cuissot,Adam Czarnołęski and Wiesław Białas - p.212 HUMAN SPECTRIN SRC HOMOLOGY 3 DOMAIN BINDING PROTEIN 1 (HSSH3BP1) IS A POTENTIAL REGULATOR OF MACROPINOCYTOSIS AND A PUTATIVE TUMOR SUPPRESSOR Leszek Kotula, Jiliu Xu and Dorota Ziemnicka - p.213 THE ROLE OF THE NON-LOCAL BENDING ENERGY IN THE RED BLOOD CELL RESPONSE TO MEMBRANE DEFORMATIONS Drago Kuzman, Marjan Gros, Sonja Vrhovec, Saša Svetina and Boštjan Žekš - p.215 INTERACTION OF SPECTRIN WITH THE LOW MOLECULAR WEIGHT PHOSPHOTYROSINE PHOSPHATASE (LMW-PTP) Marie-Christine Lecomte, GaĂ«l Nicolas, Catherine Fournier, Colette Galand, Laurence Malbert-Colas, Odile Bournier, Didier Dhermy and Bernard Grandchamp - p.216 DEFORMATION-ENHANCED FLUCTUATIONS OF THE ERYTHROCYTE'S SPECTRIN-ACTIN NODES AND RELATION TO SINGLE MOLECULE MEASURES OF SPECTRIN UNFOLDING James C-M. Lee and Dennis E. Discher - p.217 STRUCTURAL AND THERMODYNAMIC STUDIES OF CLONED FRAGMENTS OF SPECTRIN Dongning Li, Ruby I. MacDonald, Alfonso Mondragón, Edwin V. Pozharski, Julie A. Ruffatti and Valerie L. Tokars - p.218 MOTILE MEMBRANE SKELETONS: WHAT NEUTROPHILS AND MUSCLES HAVE IN COMMON Elizabeth J. Luna, Kersi N. Pestonjamasp, Thomas Nebl, Yu Chen, Cheryl Gatto, Sang W. Oh and Robert K. Pope - p.219 THE ROLE OF FOCAL ADHESION KINASE (FAK) IN CONTACT INHIBITION Joanna Miłoszewska, Halina Trembacz, Maciej Małecki and Przemysław Janik - p.220 A DYNAMIC SPECTRIN-DIMER SELF ASSOCIATION REGULATES OF RED CELL MEMBRANE MECHANICAL STABILITY Narla Mohandas, Xiu-Li An, Marie Christine Lecomte and Walter Gratzer - p.221 THE MEMBRANE SKELETON: MECHANOPROTECTOR AND MEDIATOR OF MECHANOSENSITIVE SURFACE AREA Catherine E. Morris - p.222 MOLECULAR STUDIES OF THE ERYTHROCYTE SPECTRIN TETRAMERIZATION REGION Sunghyouk Park, Shahila Mehboob, Bing-Hao Luo, Michael G. Hurtuk, M. E. Johnson and Leslie W.-M. Fung - p.224 OVEREXPRESSION OF 4.1R DISTURBS MICROTUBULE ORGANIZATION Carmen M. Pérez-Ferreiro, Carlos M. Luque, Alicia Pérez-González, Elimelec Sendino and Isabel Correas - p.225 FUNCTIONAL FLEXIBILITY OF DYSTROGLYCAN, A TRANSMEMBRANE LINKER BETWEEN THE EXTRACELLULAR MATRIX AND THE CYTOSKELETON Tamara C. Petrucci, Gianfranco Macchia, Pompeo Macioce, Andrea Brancaccio, Paola Paggi and Marina Ceccarini - p.226 CYTOSKELETAL PROTEINS IN HEART MUSCLE Jennifer C. Pinder, Pamela M. Taylor-Harris, Nandini V.L. Hayes, Philippe D.Gascard, Alison M.Maggs and Anthony J. Baines - p.227 PLANT LONG-CHAIN POLYPRENOLS AS CHEMOTAXONOMIC MARKERS Magdalena Roślińska, Krystyna Walińska, Ewa Świeżewska and Tadeusz Chojnacki - p.228 NEWER BIOPHYSICAL METHODS FOR MEMBRANE ANALYSIS IN NTACT RED CELLS Stephen B. Shohet - p.229 MECHANICAL AND FUNCTIONAL ASPECTS OF MEMBRANE SKELETONS Saša Svetina, Bojan Božič, Jure Derganc and Boštjan Žekš - p.230 FILAMENTOUS STRUCTURES IN AMOEBA PROTEUS VISUALIZED IN SCANNING MICROSCOPE Anna Wasik, Lucyna Grębecka, Andrzej Grębecki, Joanna Kołodziejczyk and Małgorzata Gołębiowska - p.231 INTERACTION OF MAJOR PHOSPHOLIPIDS OF ERYTHROCYTE MEMBRANE'S INNER LEAFLET (PS, PE) WITH PHENOTHIAZINE METHANESULFONYLAMIDES Olga Wesołowska, Andrzej B. Hendrich, Noboru Motohashi and Krystyna Michalak - p.232 DECREASE IN 2,2,6,6-TETRAMETHYL-PIPERIDINE-1-OXYL (TEMPO) EPR SIGNAL IN PEROXYNITRITE-TREATED ERYTHROCYTE MEMBRANES Anna Wróbel - p.233 CRYSTAL STRUCTURE OF THE ?-ACTININ ROD: FOUR SPECTRIN REPEATS FORMING A TIGHT DIMER Jari Jari Ylänne, Klaus Scheffzek, Paul Young and Matti Saraste - p.234 |
| Full text of the abstracts aviable | |