358 results on '"Caiafa, P."'
Search Results
202. Acid-soluble tightly-bound chromatin proteins and DNA methylation
- Author
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Reale A, Rispoli M, maria d'erme, Caiafa P, and Strom R
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DNA-Binding Proteins ,Enzyme Activation ,Humans ,DNA ,DNA Modification Methylases ,Methylation ,Chromatin - Published
- 1989
203. Association of histone-like tightly-bound proteins to hypermethylated nucleosomes
- Author
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Rispoli, M., Anna Reale, Allegra, P., Strom, R., and Caiafa, P.
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chromatin structure ,Base Composition ,Swine ,Placenta ,tightly -bound proteins ,Kidney ,Methylation ,Nucleosomes ,DNA-Binding Proteins ,Enzyme Activation ,Histones ,Pregnancy ,Animals ,Humans ,histones, tightly -bound proteins, chromatin structure ,Female ,DNA Modification Methylases - Published
- 1989
204. Distribution of tissue specific tghtly bound non-histone proteins in chromatine fractions obtained by DNAse II digestion
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Lonigro, Incoronata, Allegra, P, Altieri, F, Tommasetti, A, and Caiafa, P.
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- 1985
205. DO TIGHTLY-BOUND CHROMATIN PROTEINS PLAY A ROLE IN DNA METHYLATION
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Caiafa, P., Mastrantonio, S., Attina, M., Rispoli, M., Anna Reale, and Strom, R.
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Binding Sites ,Hydrolysis ,Placenta ,dna methylation ,DNA ,In Vitro Techniques ,Methylation ,Catalysis ,Gas Chromatography-Mass Spectrometry ,DNA-Binding Proteins ,Pregnancy ,chromatin ,nuclear matrix ,Deoxyribonuclease I ,Humans ,Female ,DNA Modification Methylases ,Chromatography, High Pressure Liquid - Abstract
When chromatin matrix, "stripped" from its loosely-bound components by extraction with 3 M NaCl, is extensively digested with DNAase I, a fraction is obtained, which carries no endogenous DNA methyltransferase activity but which is a good substrate for externally added enzyme. Under the same conditions, protein-free DNA isolated from this fraction can instead hardly be methylated, this different behaviour pointing to a role of DNA-tightly-bound proteins in favoring or promoting the catalytic action of the enzyme. A similar stimulation of enzymatic methylation could also be shown when, in the presence of this same fraction, single stranded Micrococcus luteus DNA was incubated with placental methyltransferase, using S-adenosylmethionine as a methyl donor. This finding can be correlated to the existence, in chromatin loops, of small regions which resist digestion by DNAase I also after high-salt removal of their loosely-bound components (presumably because of the presence of tightly-bound proteins) and whose DNA is characterized by high methylation levels and, at the same time, by high relative content of thymine.
- Published
- 1988
206. Distribution of tightly bound non histone proteins in chromatin fractions obtained by DNAse II digestion
- Author
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Lonigro, Incoronata, Altieri, F, Allegra, P, and Caiafa, P.
- Published
- 1985
207. Distribution of tissue-specific tightly bound non-histone proteins in chromatin fractions obtained by DNAase II digestion
- Author
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Lonigro, I. I., Allegra, P., Fabio ALTIERI, Tomassetti, A., and Caiafa, P.
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non-histone chromatin proteins ,Endodeoxyribonucleases ,Time Factors ,Chromosomal Proteins, Non-Histone ,Swine ,Antigens, Nuclear ,Kidney ,Chromatin ,Nucleoproteins ,Liver ,Solubility ,Organ Specificity ,nuclear matrix ,Animals ,chromatin ,Subcellular Fractions - Abstract
The tissue specificity of a particular class of non-histone chromatin proteins characterized by strong bonds to DNA, tightly bound non-histone chromatin proteins (t.b. NHCp), was investigated. In connection with its possible role in the cell differentiation, its distribution in chromatin fractions obtained by digestion with DNAase II and precipitation with MgCl2 was studied. It was shown by bidimensional gel electrophoresis that some proteins of this class are tissue specific for pig liver compared to kidney chromatin. For both tissues, most tissue specific proteins were found to be peculiar to the chromatin fraction that remains insoluble after prolonged digestion with DNAase II. The proteins found in this chromatin fraction are common to the nuclear matrix proteins and, considering the chromatin organization, may be involved in the attachment point for loops in the axial matrix structure. It is possible that proteins of this class, directly or indirectly, play an important role in the regulation of transcription and differentiation in eukaryotes, by modulating the supercoiled state of DNA.
- Published
- 1985
208. Non histone proteins tightly bound to loops and matrix from pig kidnei chromatin
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Lonigro, Incoronata, Tommasetti, A, and Caiafa, P.
- Published
- 1986
209. Inhibition of poly(ADP‐ribosyl)ation induces DNA hypermethylation: a possible molecular mechanism
- Author
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Zardo, Giuseppe, Reale, Anna, Passananti, Claudio, Pradhan, Sriharsa, Buontempo, Serena, De Matteis, Giovanna, Adams, Roger L.P., and Caiafa, Paola
- Abstract
The pattern of DNA methylation established during embryonic development is necessary for the control of gene expression and is preserved during the replicative process. DNA regions of about 1–2 kb in size, termed CpG islands and located mostly in the promoter regions of housekeeping genes, are protected from methylation, despite being about 6–10 times richer in the dinucleotide CpG than the rest of DNA. Their unmethylated state guarantees the expression of the corresponding housekeeping genes. At present, the mechanism by which CpG islands remain protected from methylation is not clear. However, some results suggest that poly(ADPribosyl)ation, an enzymatic process that introduces a postsynthetic modification onto chromatin proteins, might be involved. Here we show in L929 mouse fibroblast cells that inhibition of poly(ADP‐ribose) polymerase(s) at different cell‐cycle phases increases the mRNA and protein levels of the major maintenance DNA methyltransferase (DNMT1) in G1/S border. Increase of DNMT1 results in a premature PCNA‐DNMT1 complex formation, which facilitates robust maintenance, as well as de novoDNA methylation processes during the G1/S border, which leads to abnormal hypermethylation.
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- 2002
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210. Effect of polyenic antibiotics on Ehrlich ascites and Novikoff hepatoma cells
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Mondovì, B., Strom, R., Finazzi Agrò, A., Caiafa, P., Sole, P., Argante BOZZI, Rotilio, G., and Rossi Fanelli, A.
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DNA Replication ,Male ,Antifungal Agents ,Carcinoma, Hepatocellular ,Cell Membrane Permeability ,polyenic antibiotics ,Tritium ,Cell Line ,Lactones ,Mice ,Oxygen Consumption ,Glutamates ,Cell Movement ,Culture Techniques ,Strophanthins ,Chromium Isotopes ,Animals ,Carcinoma, Ehrlich Tumor ,Cell Nucleus ,Carbon Isotopes ,Antibiotics, Antineoplastic ,hepatoma ,neoplasia ,Cell Membrane ,Fatty Acids ,Liver Neoplasms ,DNA, Neoplasm ,Neoplasms, Experimental ,Trypan Blue ,Hydrogen-Ion Concentration ,Liver Regeneration ,Rats ,Potassium ,Thymidine
211. IDENTIFICATION OF NON HISTONE CHROMATIN PROTEINS TIGHTLY BOUND TO NUCLEOSOMES
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Caiafa, P., primary, Cioffari, M. R. Scarpati, additional, Altieri, F., additional, Allegra, P., additional, and Lonigro, R., additional
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- 1981
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212. Succinate dehydrogenase II. The effect of phospholipases on particulate and soluble succinate dehydrogenase
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Cerletti, P., primary, Caiafa, P., additional, Giordano, M.G., additional, and Giovenco, M.A., additional
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- 1969
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213. TET1 Promotes Leukemic Growth in T-ALL Via Maintenance of 5-Hydroxymethylation Marks and Can be Antagonized By the PARP Inhibitor Olaparib
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Sahin, Deniz, Bamezai, Shiva, Ciccarone, Fabio, Fischbein, Elena, Mohr, Fabian, Mulaw, Medhanie A., Pulikkottil Jose, Alex, Caiafa, Paola, Meyer, Lüder-Hinrich, Debatin, Klaus-Michael, Kronnie, Geertruy te, Döhner, Konstanze, Döhner, Hartmut, Feuring-Buske, Michaela, Buske, Christian, and Rawat, Vijay P.S.
- Abstract
Mulaw: NuGEN: Honoraria. Buske:Celltrion, Inc.: Consultancy, Honoraria.
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- 2016
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214. The PARP Inhibitor Olaparib Antagonizes Leukemic Growth Induced By TET1 Overexpression in AML1-ETO Positive Acute Myeloid Leukemia
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Bamezai, Shiva, He, Jing, Sahin, Deniz, Mohr, Fabian, Ciccarone, Fabio, Vegi, Naidu M, Pulikkottil Jose, Alex, Mulaw, Medhanie A., Caiafa, Paola, Döhner, Konstanze, Döhner, Hartmut, Feuring-Buske, Michaela, Buske, Christian, and Rawat, Vijay P.S.
- Abstract
Mulaw: NuGEN: Honoraria. Buske:Celltrion, Inc.: Consultancy, Honoraria.
- Published
- 2016
- Full Text
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215. The spontaneous dwarf rat (SDR) has selectively reduced apoptosis and proliferation in response to AOM
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Collins, Jason, Choi, Young, Marchandani, Jai, Caiafa, Guy, Swanson, Stephen, and Carroll, Robert
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- 2003
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216. Pet Dental Health Week breaks records
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Caiafa, Dr Anthony
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- 2002
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217. CORRESPONDENCE
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CAIAFA, AM
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- 2000
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218. Does hypomethylation of linker DNA play a role in chromatin condensation?
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Caiafa, P., Reale, A., Santoro, R., and D'Erme, M.
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- 1995
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219. Ageing affects subtelomeric DNA methylation in blood cells from a large European population enrolled in the MARK-AGE study
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Michele Zampieri, Beatrix Grubeck-Loebenstein, Alexander Bürkle, Mikko Hurme, P. Eline Slagboom, Florence Debacq-Chainiaux, Olivier Toussaint, Maria A. Blasco, Tilman Grune, Claudio Franceschi, Ewa Sikora, Maria Giulia Bacalini, Antti Hervonen, Eugène H.J.M. Jansen, Stefano Tagliatesta, Valentina Aversano, Anna Reale, Martijn E.T. Dollé, Miriam Capri, Marco Malavolta, Maria Moreno-Villanueva, Efstathios S. Gonos, Christiane Schön, Nicolle Breusing, Fabio Ciccarone, Jürgen Bernhardt, Paola Caiafa, Bacalini M.G., Reale A., Malavolta M., Ciccarone F., Moreno-Villanueva M., Dolle M.E.T., Jansen E., Grune T., Gonos E.S., Schon C., Bernhardt J., Grubeck-Loebenstein B., Sikora E., Toussaint O., Debacq-Chainiaux F., Capri M., Hervonen A., Hurme M., Slagboom P.E., Breusing N., Aversano V., Tagliatesta S., Franceschi C., Blasco M.A., Burkle A., Caiafa P., and Zampieri M.
- Subjects
0301 basic medicine ,Male ,Aging ,Offspring ,ageing ,subtelomere ,epigenetics ,DNA methylation ,Down syndrome ,centenarian offspring ,Population ,Context (language use) ,Centenarian offspring ,Biology ,03 medical and health sciences ,0302 clinical medicine ,ddc:570 ,Blood Cell ,Humans ,Epigenetics ,Settore BIO/10 ,education ,Cross-Sectional Studie ,Genetics ,Aged, 80 and over ,education.field_of_study ,Subtelomere ,Blood Cells ,Epigenetic ,Methylation ,Europe ,Ageing ,030104 developmental biology ,Cross-Sectional Studies ,030220 oncology & carcinogenesis ,Female ,Original Article ,Geriatrics and Gerontology ,Human - Abstract
Ageing leaves characteristic traces in the DNA methylation make-up of the genome. However, the importance of DNA methylation in ageing remains unclear. The study of subtelomeric regions could give promising insights into this issue. Previously reported associations between susceptibility to age-related diseases and epigenetic instability at subtelomeres suggest that the DNA methylation profile of subtelomeres undergoes remodelling during ageing. In the present work, this hypothesis has been tested in the context of the European large-scale project MARK-AGE. In this cross-sectional study, we profiled the DNA methylation of chromosomes 5 and 21 subtelomeres, in more than 2000 age-stratified women and men recruited in eight European countries. The study included individuals from the general population as well as the offspring of nonagenarians and Down syndrome subjects, who served as putative models of delayed and accelerated ageing, respectively. Significant linear changes of subtelomeric DNA methylation with increasing age were detected in the general population, indicating that subtelomeric DNA methylation changes are typical signs of ageing. Data also show that, compared to the general population, the dynamics of age-related DNA methylation changes are attenuated in the offspring of centenarian, while they accelerate in Down syndrome individuals. This result suggests that subtelomeric DNA methylation changes reflect the rate of ageing progression. We next attempted to trace the age-related changes of subtelomeric methylation back to the influence of diverse variables associated with methylation variations in the population, including demographics, dietary/health habits and clinical parameters. Results indicate that the effects of age on subtelomeric DNA methylation are mostly independent of all other variables evaluated. Supplementary Information The online version contains supplementary material available at 10.1007/s11357-021-00347-9.
- Published
- 2021
220. TET2 gene expression and 5-hydroxymethylcytosine level in multiple sclerosis peripheral blood cells
- Author
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Paola Caiafa, Elisabetta Valentini, Maria Giulia Bacalini, Viviana Annibali, Fabio Ciccarone, Michele Zampieri, Vito A. G. Ricigliano, Roberta Calabrese, Claudio Franceschi, Rosella Mechelli, Marco Salvetti, Tiziana Guastafierro, Department of Cellular Biotechnologies and Hematology, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP), Department of Experimental Pathology, Alma Mater Studiorum Università di Bologna [Bologna] (UNIBO), Center for Experimental Neurological Therapies (CENTERS), Neurology and Department of Neuroscience, Mental Health and Sensory Organs, Neuroimmunology Unit, Clinical and Behavioral Neurology - Neuroscienze e riabilitazione, IRCCS Fondazione Santa Lucia [Roma], This work was supported by FISM (Fondazione Italiana SclerosiMultipla) Cod. 2011/R/9'., Calabrese, R., Valentini, E., Ciccarone, F., Guastafierro, T., Bacalini, M.G., Ricigliano, V.A.G., Zampieri, M., Annibali, V., Mechelli, R., Franceschi, C., Salvetti, M., and Caiafa, P.
- Subjects
Adult ,DNA (Cytosine-5-)-Methyltransferase 1 ,Male ,DNA-Binding Protein ,Down-Regulation ,Gene Expression ,Ten-Eleven Translocation 2 ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Biology ,Dioxygenases ,Multiple sclerosis ,chemistry.chemical_compound ,Cytosine ,Proto-Oncogene Proteins ,5-Hydroxymethylcytosine ,Multiple Sclerosi ,Gene expression ,Humans ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Epigenetics ,DNA (Cytosine-5-)-Methyltransferases ,Settore BIO/10 ,Promoter Regions, Genetic ,Molecular Biology ,Epigenomics ,Proto-Oncogene Protein ,DNA methylation ,epigenetics ,5-hydroxymethylcytosine ,multiple sclerosis ,dna methylation ,peripheral blood mononuclear cells ,ten-eleven translocation 2 ,Epigenetic ,Promoter ,Peripheral blood mononuclear cell ,Molecular biology ,DNA-Binding Proteins ,DNA demethylation ,chemistry ,DNA (Cytosine-5-)-Methyltransferase ,Case-Control Studies ,Peripheral blood mononuclear cells ,DNMT1 ,5-Methylcytosine ,Leukocytes, Mononuclear ,Molecular Medicine ,Female ,Case-Control Studie ,Human - Abstract
International audience; Aberrant DNA methylation can lead to genome destabilization and to deregulated gene expression. Recently, 5-hydroxymethylcytosine (5hmC), derived from oxidation of 5-methylcytosine (5mC) by the Ten-Eleven Translo-cation (TET) enzymes, has been detected. 5hmC is now considered as a new epigenetic DNA modification with relevant roles in cell homeostasis regulating DNA demethylation and transcription. Our aim was to investigate possible changes in the DNA methylation/demethylation machinery in MS. We assessed the expression of enzymes involved in DNA methylation/demethylation in peripheral blood mononuclear cells (PBMCs) from 40 subjects with MS and 40 matched healthy controls. We performed also, DNA methylation analysis of specific promoters and analysis of global levels of 5mC and 5hmC. We show that TET2 and DNMT1 expression is significantly down-regulated in MS PBMCs and it is associated with aberrant methylation of their promoters. Furthermore , 5hmC is decreased in MS PBMCs, probably as a result of the diminished TET2 level.
- Published
- 2014
221. Validation of suitable internal control genes for expression studies in aging
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Fabio Ciccarone, Michele Zampieri, Roberta Calabrese, Paola Caiafa, Marta Chevanne, Maria Giulia Bacalini, Maria Moreno-Villanueva, Anna Reale, Alexander Bürkle, Tiziana Guastafierro, Zampieri, M., Ciccarone, F., Guastafierro, T., Bacalini, M.G., Calabrese, R., Moreno-Villanueva, M., Reale, A., Chevanne, M., Bürkle, A., and Caiafa, P.
- Subjects
Senescence ,Adult ,Aging ,Hypoxanthine Phosphoribosyltransferase ,ageing ,real-time PCR ,reference genes selection ,Reproducibility of Result ,Gene Expression ,Biology ,Gene ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,law ,Transcription (biology) ,ddc:570 ,Reference genes ,Gene expression ,Humans ,RNA, Messenger ,Settore BIO/10 ,Housekeeping gene ,Polymerase chain reaction ,030304 developmental biology ,Aged ,Genetics ,0303 health sciences ,Housekeeping genes ,Reverse Transcriptase Polymerase Chain Reaction ,Confounding ,PBMC ,Reproducibility of Results ,Reference Standards ,Middle Aged ,Real-time PCR normalization ,Algorithm ,Genes ,GUSB ,Leukocytes, Mononuclear ,Reference Standard ,Algorithms ,030217 neurology & neurosurgery ,Human ,Developmental Biology - Abstract
Quantitative data from experiments of gene expression are often normalized through levels of housekeeping genes transcription by assuming that expression of these genes is highly uniform. This practice is being questioned as it becomes increasingly clear that the level of housekeeping genes expression may vary considerably in certain biological samples. To date, the validation of reference genes in aging has received little attention and suitable reference genes have not yet been defined. Our aim was to evaluate the expression stability of frequently used reference genes in human peripheral blood mononuclear cells with respect to aging. Using quantitative RT-PCR, we carried out an extensive evaluation of five housekeeping genes, i.e. 18s rRNA, ACTB, GAPDH, HPRT1 and GUSB, for stability of expression in samples from donors in the age range 35-74 years. The consistency in the expression stability was quantified on the basis of the coefficient of variation and two algorithms termed geNorm and NormFinder. Our results indicated GUSB be the most suitable transcript and 18s the least for accurate normalization in PBMCs. We also demonstrated that aging is a confounding factor with respect to stability of 18s, HPRT1 and ACTB expression, which were particularly prone to variability in aged donors. © 2009 Elsevier Ireland Ltd.
- Published
- 2010
222. Inhibition of Poly(ADP-Ribosyl)ation Allows DNA Hypermethylation
- Author
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Giuseppe Zardo, Paola Caiafa, Maria Malanga, Anna Reale, Jordanka Zlatanova, University of Zurich, Szyf, M, M. Szyf, Reale, A, Zardo, G, Malanga, Maria, Zlatanova, J, and Caiafa, P.
- Subjects
DNA methylation ,biology ,Chemistry ,poly(ADP-ribosyl)ation ,Promoter ,10079 Institute of Veterinary Pharmacology and Toxicology ,Methylation ,Chromatin ,Cell biology ,chemistry.chemical_compound ,genomic DNA ,Histone ,CpG site ,biology.protein ,570 Life sciences ,DNA - Abstract
This chapter emphasizes that along the chain of events that induce DNA methylation-dependent chromatin condensation, a post-synthetic modification other than histone acetylation, poly(ADP-ribosyl)ation, participates in the establishment and maintenance of methylation-free regions of chromatin. In fact, several lines of in vitro and in vivo evidence have shown that poly(ADP-ribosyl)ation is involved in the control of DNA methylation pattern, protecting genomic DNA from full methylation. More recent studies have provided some clues to the understanding of the molecular mechanism(s) connecting poly(ADP-ribosyl)ation with DNA methylation. We aim here to demonstrate the direct correlation existing between inhibition of poly(ADP-ribose) polymerases and DNA hypermethylation, and to describe some possible mechanisms underlying this molecular link. We will then present our hypothesis that the inhibition of the poly(ADP-ribosyl)ation process in the cell may be responsible for the anomalous hypermethylation of oncosuppressor gene promoters during tumorigenesis and to suggest the possibility that an active poly(ADP-ribosyl)ation process is also involved in maintaining the unmethylated state of CpG islands in normal cells.
- Published
- 2007
223. Gli strumenti di reporting sociale negli enti locali: a che punto siamo?
- Author
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SIBONI, BENEDETTA, A. ALLINI, B. ANTONELLI, S. ASTOLFI, L. BIONDI, A.M. BISCOTTI, A. BONFANTI, L. BOSETTI, G. BOSSO, L. BROGONZOLI, F. BRUNO, R. CAIAFA, P. CARENZO, A. CASALENA, R. CASSANO, P. CASTELLANI, C. CORSI, E. COSTA, E. CRESTI, M. DE NICOLA, C. DEL GESSO, A. DELLA PORTA, D. DI BERARDINO, T. DI CIMBRINI, P. DI GREGORIO, F. FACCIA, G. FALDETTA, D. FELAPPI, C. FERRONE, S. GIGLI, S. GIULIANI, C. GULLUSCIO, G. GUZZO, L. IANNI, B. IANNONE, P. LA MARCA, F. LA ROSA, I. LUPERTO, F. MANDANICI, R. MEDEI, M.C. MONTAGANO, F. MONTEDURO, C PARISI, A. PASSARO, M. PEDRINI, A. PETTINELLA, F. PORTADIBASSO, M. PUZZO, A. QUINTILIANI, G. RAPPA, R. REINA, C. ROMANO, C. ROSSATO, R. RUSCIANO, F. SANSALVADORE, B. SIBONI, L. TAMPIERI, P. VOLA, M. ZIFARO, and B. Siboni
- Subjects
STAKEHOLDERS ,RENDICONTAZIONE SOCIALE ,ENTI LOCALI ,ACCOUNTABILITY ,BILANCIO SOCIALE - Abstract
Il lavoro analizza le ragioni che hanno portato alla diffusione della rendicontazione sociale negli enti locali italiani e presenta alcuni primi risultati di una indagine svolta dalla scrivente per rilevarne il grado di diffusione e le caratteristiche.
- Published
- 2007
224. Poly(ADP-ribosyl)ation system in nuclear matrix associated regions of rat testis chromatin
- Author
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P. QUESADA, M. D'ERME, L. ATORINO, FARAONE MENNELLA, MARIA ROSARIA, P. CAIAFA, FARINA, BENEDETTA, Quesada, PIERINA MARIA, D'Erme, M, Atorino, L, FARAONE MENNELLA, MARIA ROSARIA, Caiafa, P, Farina, B., P., Quesada, M., D'Erme, L., Atorino, P., Caiafa, and Farina, Benedetta
- Published
- 1995
225. DNA Methylation Analysis of Ribosomal DNA in Adults With Down Syndrome.
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Ravaioli F, Zampieri M, Morandi L, Pirazzini C, Pellegrini C, De Fanti S, Gensous N, Pirazzoli GL, Sambati L, Ghezzo A, Ciccarone F, Reale A, Monti D, Salvioli S, Caiafa P, Capri M, Bürkle A, Moreno-Villanueva M, Garagnani P, Franceschi C, and Bacalini MG
- Abstract
Control of ribosome biogenesis is a critical aspect of the regulation of cell metabolism. As ribosomal genes (rDNA) are organized in repeated clusters on chromosomes 13, 14, 15, 21, and 22, trisomy of chromosome 21 confers an excess of rDNA copies to persons with Down syndrome (DS). Previous studies showed an alteration of ribosome biogenesis in children with DS, but the epigenetic regulation of rDNA genes has not been investigated in adults with DS so far. In this study, we used a targeted deep-sequencing approach to measure DNA methylation (DNAm) of rDNA units in whole blood from 69 adults with DS and 95 euploid controls. We further evaluated the expression of the precursor of ribosomal RNAs (RNA45S) in peripheral blood mononuclear cells (PBMCs) from the same subjects. We found that the rDNA promoter tends to be hypermethylated in DS concerning the control group. The analysis of epihaplotypes (the combination of methylated and unmethylated CpG sites along the same DNA molecule) showed a significantly lower intra-individual diversity in the DS group, which at the same time was characterized by a higher interindividual variability. Finally, we showed that RNA45S expression is lower in adults with DS. Collectively, our results suggest a rearrangement of the epigenetic profile of rDNA in DS, possibly to compensate for the extranumerary rDNA copies. Future studies should assess whether the regulation of ribosome biogenesis can contribute to the pathogenesis of DS and explain the clinical heterogeneity characteristic of the syndrome., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer AT declared a shared affiliation with the authors MZ, AR, and PC to the handling editor at the time of review., (Copyright © 2022 Ravaioli, Zampieri, Morandi, Pirazzini, Pellegrini, De Fanti, Gensous, Pirazzoli, Sambati, Ghezzo, Ciccarone, Reale, Monti, Salvioli, Caiafa, Capri, Bürkle, Moreno-Villanueva, Garagnani, Franceschi and Bacalini.)
- Published
- 2022
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226. Nutritional Factors Modulating Alu Methylation in an Italian Sample from The Mark-Age Study Including Offspring of Healthy Nonagenarians.
- Author
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Giacconi R, Malavolta M, Bürkle A, Moreno-Villanueva M, Franceschi C, Capri M, Slagboom PE, Jansen EHJM, Dollé MET, Grune T, Weber D, Hervonen A, Stuetz W, Breusing N, Ciccarone F, Zampieri M, Aversano V, Caiafa P, Formentini L, Piacenza F, Pierpaoli E, Basso A, Provinciali M, and Cardelli M
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- Adult, Aged, Aging blood, Antioxidants analysis, CpG Islands, Diet, Female, Healthy Volunteers, Humans, Italy, Lipoproteins blood, Lipoproteins genetics, Longevity genetics, Male, Middle Aged, Nutritional Status, Trace Elements blood, Aging genetics, Alu Elements, DNA Methylation, Nutrients blood
- Abstract
Alu hypomethylation promotes genomic instability and is associated with aging and age-related diseases. Dietary factors affect global DNA methylation, leading to changes in genomic stability and gene expression with an impact on longevity and the risk of disease. This preliminary study aims to investigate the relationship between nutritional factors, such as circulating trace elements, lipids and antioxidants, and Alu methylation in elderly subjects and offspring of healthy nonagenarians. Alu DNA methylation was analyzed in sixty RASIG (randomly recruited age-stratified individuals from the general population) and thirty-two GO (GeHA offspring) enrolled in Italy in the framework of the MARK-AGE project. Factor analysis revealed a different clustering between Alu CpG1 and the other CpG sites. RASIG over 65 years showed lower Alu CpG1 methylation than those of GO subjects in the same age class. Moreover, Alu CpG1 methylation was associated with fruit and whole-grain bread consumption, LDL2-Cholesterol and plasma copper. The preserved Alu methylation status in GO, suggests Alu epigenetic changes as a potential marker of aging. Our preliminary investigation shows that Alu methylation may be affected by food rich in fibers and antioxidants, or circulating LDL subfractions and plasma copper.
- Published
- 2019
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227. Histone Modifications Drive Aberrant Notch3 Expression/Activity and Growth in T-ALL.
- Author
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Tottone L, Zhdanovskaya N, Carmona Pestaña Á, Zampieri M, Simeoni F, Lazzari S, Ruocco V, Pelullo M, Caiafa P, Felli MP, Checquolo S, Bellavia D, Talora C, Screpanti I, and Palermo R
- Abstract
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive blood cancer caused by the deregulation of key T-cell developmental pathways, including Notch signaling. Aberrant Notch signaling in T-ALL occurs by NOTCH1 gain-of-function mutations and by NOTCH3 overexpression. Although NOTCH3 is assumed as a Notch1 target, machinery driving its transcription in T-ALL is undefined in leukemia subsets lacking Notch1 activation. Here, we found that the binding of the intracellular Notch3 domain, as well as of the activated Notch1 fragment, to the NOTCH3 gene locus led to the recruitment of the H3K27 modifiers JMJD3 and p300, and it was required to preserve transcriptional permissive/active H3K27 marks and to sustain NOTCH3 gene expression levels. Consistently, pharmacological inhibition of JMJD3 by GSKJ4 treatment or of p300 by A-485 decreased the levels of expression of NOTCH3, NOTCH1 and of the Notch target genes DELTEX1 and c-Myc and abrogated cell viability in both Notch1- and Notch3-dependent T-cell contexts. Notably, re-introduction of exogenous Notch1, Notch3 as well as c-Myc partially rescued cells from anti-growth effects induced by either treatment. Overall our findings indicate JMJD3 and p300 as general Notch1 and Notch3 signaling co-activators in T-ALL and suggest further investigation on the potential therapeutic anti-leukemic efficacy of their enzymatic inhibition in Notch/c-Myc axis-related cancers and diseases.
- Published
- 2019
- Full Text
- View/download PDF
228. DNA methylation dynamics in aging: how far are we from understanding the mechanisms?
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Ciccarone F, Tagliatesta S, Caiafa P, and Zampieri M
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- Animals, Chromatin genetics, Histones genetics, Humans, Aging physiology, Chromatin metabolism, DNA Methylation physiology, Epigenesis, Genetic physiology, Histones metabolism, Protein Processing, Post-Translational physiology
- Abstract
DNA methylation is currently the most promising molecular marker for monitoring aging and predicting life expectancy. However, the mechanisms underlying age-related DNA methylation changes remain mostly undiscovered. Here we discuss the current knowledge of the dynamic nature of DNA epigenome landscape in mammals, and propose putative molecular mechanisms for aging-associated DNA epigenetic changes. Specifically, we describe age-related variations of methylcytosine and its oxidative derivatives in relation to the dynamics of chromatin structure, histone post-translational modifications and their modulators. Finally, we are proposing a conceptual framework that could explain the complex nature of the effects of age on DNA methylation patterns. This combines the accumulation of DNA methylation noise and also all of the predictable, site-specific DNA methylation changes. Gathering information in this area would pave the way for future investigation aimed at establishing a possible causative role of epigenetic mechanisms in aging., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2018
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229. Zinc-Induced Metallothionein in Centenarian Offspring From a Large European Population: The MARK-AGE Project.
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Giacconi R, Costarelli L, Piacenza F, Basso A, Bürkle A, Moreno-Villanueva M, Grune T, Weber D, Stuetz W, Gonos ES, Schön C, Grubeck-Loebenstein B, Sikora E, Toussaint O, Debacq-Chainiaux F, Franceschi C, Hervonen A, Slagboom E, Ciccarone F, Zampieri M, Caiafa P, Jansen E, Dollé MET, Breusing N, Mocchegiani E, and Malavolta M
- Subjects
- Adult, Aged, Aged, 80 and over, Cell Culture Techniques, Cross-Sectional Studies, Europe, Female, Flow Cytometry, Humans, Male, Middle Aged, Oxidative Stress, RNA metabolism, Real-Time Polymerase Chain Reaction, Biomarkers metabolism, Leukocytes, Mononuclear metabolism, Metallothionein metabolism, Zinc pharmacology
- Abstract
Metallothionein (MT) family are cysteine-rich proteins that regulate zinc (Zn) homeostasis and protect against oxidative damage. Studies in transgenic mice have shown that MT favorably influence longevity, although their role in human aging is not completely understood. Within the European multicenter study MARK-AGE, we analyzed MT induction after Zn treatment in peripheral blood mononuclear cells (PBMCs) and its relation with redox biomarkers in 2,936 age-stratified subjects (35-75 years) including the general population (RASIG), centenarian offspring (GO), and their spouses (SGO). We found that the lymphocyte capability to induce MT in response to Zn is not affected by aging. However, GO participants showed lower Zn-induced MT and increased basal expression of MT1A, MT1X, and ZnT-1 genes than RASIG subjects. Moreover, Zn-induced MT levels were found to be inversely related with oxidative stress markers (plasma protein carbonyls, 3-nitrotyrosine, and malondialdehyde) in the whole population, but not in GO subjects. In conclusion, our results support the hypothesis that the response to Zn is attenuated in PBMCs of centenarian offspring compared to the general population as a consequence of a tighter control of Zn homeostasis which is likely to provide them constant protection against stress stimuli over the whole lifespan.
- Published
- 2018
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230. DNA Hydroxymethylation Levels Are Altered in Blood Cells From Down Syndrome Persons Enrolled in the MARK-AGE Project.
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Ciccarone F, Valentini E, Malavolta M, Zampieri M, Bacalini MG, Calabrese R, Guastafierro T, Reale A, Franceschi C, Capri M, Breusing N, Grune T, Moreno-Villanueva M, Bürkle A, and Caiafa P
- Subjects
- 5-Methylcytosine analogs & derivatives, 5-Methylcytosine blood, Adult, Aged, Cross-Sectional Studies, Epigenesis, Genetic, Europe, Female, Humans, Immunoblotting, Italy, Male, Middle Aged, Mixed Function Oxygenases blood, Proto-Oncogene Proteins blood, RNA, Messenger blood, Aging blood, Aging genetics, DNA Methylation, Down Syndrome blood, Down Syndrome genetics, Leukocytes, Mononuclear metabolism
- Abstract
Down syndrome (DS) is caused by the presence of part or an entire extra copy of chromosome 21, a phenomenon that can cause a wide spectrum of clinically defined phenotypes of the disease. Most of the clinical signs of DS are typical of the aging process including dysregulation of immune system. Beyond the causative genetic defect, DS persons display epigenetic alterations, particularly aberrant DNA methylation patterns that can contribute to the heterogeneity of the disease. In the present work, we investigated the levels of 5-hydroxymethylcytosine and of the Ten-eleven translocation dioxygenase enzymes, which are involved in DNA demethylation processes and are often deregulated in pathological conditions as well as in aging. Analyses were carried out on peripheral blood mononuclear cells of DS volunteers enrolled in the context of the MARK-AGE study, a large-scale cross-sectional population study with subjects representing the general population in eight European countries. We observed a decrease in 5-hydroxymethylcytosine, TET1, and other components of the DNA methylation/demethylation machinery in DS subjects, indicating that aberrant DNA methylation patterns in DS, which may have consequences on the transcriptional status of immune cells, may be due to a global disturbance of methylation control in DS.
- Published
- 2018
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231. PARP1 orchestrates epigenetic events setting up chromatin domains.
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Ciccarone F, Zampieri M, and Caiafa P
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- ADP-Ribosylation, Animals, Humans, Models, Biological, Chromatin metabolism, Epigenesis, Genetic, Poly(ADP-ribose) Polymerases metabolism
- Abstract
Epigenetic events include reversible modifications of DNA and histone tails driving chromatin organization and thus transcription. The epigenetic regulation is a highly integrated process underlying the plasticity of the genomic information both in the context of complex physiological and pathological processes. The global regulatory aspects of epigenetic events are largely unknown. PARylation and PARP1 are recently emerging as multi-level regulatory effectors that modulate the topology of chromatin by orchestrating very different processes. This review focuses in particular on the role of PARP1 in epigenetics, trying to build a comprehensive perspective of its involvement in the regulation of epigenetic modifications of histones and DNA, contextualizing it in the global organization of chromatin domains in the nucleus., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2017
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232. Analysis of the machinery and intermediates of the 5hmC-mediated DNA demethylation pathway in aging on samples from the MARK-AGE Study.
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Valentini E, Zampieri M, Malavolta M, Bacalini MG, Calabrese R, Guastafierro T, Reale A, Franceschi C, Hervonen A, Koller B, Bernhardt J, Slagboom PE, Toussaint O, Sikora E, Gonos ES, Breusing N, Grune T, Jansen E, Dollé ME, Moreno-Villanueva M, Sindlinger T, Bürkle A, Ciccarone F, and Caiafa P
- Subjects
- Adult, Aged, Aging metabolism, Dioxygenases metabolism, Female, Gene Expression, Humans, Leukocytes, Mononuclear metabolism, Male, Middle Aged, Mixed Function Oxygenases metabolism, Proto-Oncogene Proteins metabolism, 5-Methylcytosine metabolism, Aging genetics, DNA Methylation, Dioxygenases genetics, Gene Expression Regulation, Mixed Function Oxygenases genetics, Proto-Oncogene Proteins genetics
- Abstract
Gradual changes in the DNA methylation landscape occur throughout aging virtually in all human tissues. A widespread reduction of 5-methylcytosine (5mC), associated with highly reproducible site-specific hypermethylation, characterizes the genome in aging. Therefore, an equilibrium seems to exist between general and directional deregulating events concerning DNA methylation controllers, which may underpin the age-related epigenetic changes. In this context, 5mC-hydroxylases (TET enzymes) are new potential players. In fact, TETs catalyze the stepwise oxidation of 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC), driving the DNA demethylation process based on thymine DNA glycosylase (TDG)-mediated DNA repair pathway. The present paper reports the expression of DNA hydroxymethylation components, the levels of 5hmC and of its derivatives in peripheral blood mononuclear cells of age-stratified donors recruited in several European countries in the context of the EU Project 'MARK-AGE'. The results provide evidence for an age-related decline of TET1 , TET3 and TDG gene expression along with a decrease of 5hmC and an accumulation of 5caC. These associations were independent of confounding variables, including recruitment center, gender and leukocyte composition. The observed impairment of 5hmC-mediated DNA demethylation pathway in blood cells may lead to aberrant transcriptional programs in the elderly., Competing Interests: All authors declare no conflict of interest.
- Published
- 2016
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233. Age-dependent expression of DNMT1 and DNMT3B in PBMCs from a large European population enrolled in the MARK-AGE study.
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Ciccarone F, Malavolta M, Calabrese R, Guastafierro T, Bacalini MG, Reale A, Franceschi C, Capri M, Hervonen A, Hurme M, Grubeck-Loebenstein B, Koller B, Bernhardt J, Schӧn C, Slagboom PE, Toussaint O, Sikora E, Gonos ES, Breusing N, Grune T, Jansen E, Dollé M, Moreno-Villanueva M, Sindlinger T, Bürkle A, Zampieri M, and Caiafa P
- Subjects
- Adult, Aged, Body Mass Index, DNA (Cytosine-5-)-Methyltransferase 1, DNA (Cytosine-5-)-Methyltransferases metabolism, Female, Gene Ontology, Humans, Life Style, Male, Middle Aged, RNA, Messenger genetics, RNA, Messenger metabolism, Regression Analysis, Risk Factors, DNA Methyltransferase 3B, Aging genetics, DNA (Cytosine-5-)-Methyltransferases genetics, Gene Expression Regulation, Developmental, Gene Expression Regulation, Enzymologic, Leukocytes, Mononuclear enzymology, White People genetics
- Abstract
Aging is associated with alterations in the content and patterns of DNA methylation virtually throughout the entire human lifespan. Reasons for these variations are not well understood. However, several lines of evidence suggest that the epigenetic instability in aging may be traced back to the alteration of the expression of DNA methyltransferases. Here, the association of the expression of DNA methyltransferases DNMT1 and DNMT3B with age has been analysed in the context of the MARK-AGE study, a large-scale cross-sectional study of the European general population. Using peripheral blood mononuclear cells, we assessed the variation of DNMT1 and DNMT3B gene expression in more than two thousand age-stratified women and men (35-75 years) recruited across eight European countries. Significant age-related changes were detected for both transcripts. The level of DNMT1 gradually dropped with aging but this was only observed up to the age of 64 years. By contrast, the expression of DNMT3B decreased linearly with increasing age and this association was particularly evident in females. We next attempted to trace the age-related changes of both transcripts to the influence of different variables that have an impact on changes of their expression in the population, including demographics, dietary and health habits, and clinical parameters. Our results indicate that age affects the expression of DNMT1 and DNMT3B as an almost independent variable in respect of all other variables evaluated., (© 2016 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.)
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- 2016
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234. Poly(ADP-Ribosyl)ation Affects Histone Acetylation and Transcription.
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Verdone L, La Fortezza M, Ciccarone F, Caiafa P, Zampieri M, and Caserta M
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- Acetylation, Animals, Benzimidazoles pharmacology, DNA (Cytosine-5-)-Methyltransferase 1, DNA (Cytosine-5-)-Methyltransferases biosynthesis, E1A-Associated p300 Protein biosynthesis, Genomic Instability, Mice, NIH 3T3 Cells, Phenanthrenes pharmacology, Tumor Necrosis Factor-alpha biosynthesis, p300-CBP Transcription Factors biosynthesis, Histones metabolism, Poly Adenosine Diphosphate Ribose metabolism, Poly(ADP-ribose) Polymerases metabolism, Transcription, Genetic
- Abstract
Poly(ADP-ribosyl)ation (PARylation) is a posttranslational protein modification catalyzed by members of the poly(ADP-ribose) polymerase (PARP) enzyme family. PARylation regulates a wide variety of biological processes in most eukaryotic cells including energy metabolism and cell death, maintenance of genomic stability, chromatin structure and transcription. Inside the nucleus, cross-talk between PARylation and other epigenetic modifications, such as DNA and histone methylation, was already described. In the present work, using PJ34 or ABT888 to inhibit PARP activity or over-expressing poly(ADP-ribose) glycohydrolase (PARG), we show decrease of global histone H3 and H4 acetylation. This effect is accompanied by a reduction of the steady state mRNA level of p300, Pcaf, and Tnfα, but not of Dnmt1. Chromatin immunoprecipitation (ChIP) analyses, performed at the level of the Transcription Start Site (TSS) of these four genes, reveal that changes in histone acetylation are specific for each promoter. Finally, we demonstrate an increase of global deacetylase activity in nuclear extracts from cells treated with PJ34, whereas global acetyltransferase activity is not affected, suggesting a role for PARP in the inhibition of histone deacetylases. Taken together, these results show an important link between PARylation and histone acetylation regulated transcription.
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- 2015
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235. Reconfiguration of DNA methylation in aging.
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Zampieri M, Ciccarone F, Calabrese R, Franceschi C, Bürkle A, and Caiafa P
- Subjects
- Aging genetics, Animals, Humans, Aging metabolism, CpG Islands physiology, DNA Methylation physiology, Epigenesis, Genetic physiology, Genome, Human physiology
- Abstract
A complex interplay between multiple biological effects shapes the aging process. The advent of genome-wide quantitative approaches in the epigenetic field has highlighted the effective impact of epigenetic deregulation, particularly of DNA methylation, on aging. Age-associated alterations in DNA methylation are commonly grouped in the phenomenon known as "epigenetic drift" which is characterized by gradual extensive demethylation of genome and hypermethylation of a number of promoter-associated CpG islands. Surprisingly, specific DNA regions show directional epigenetic changes in aged individuals suggesting the importance of these events for the aging process. However, the epigenetic information obtained until now in aging needs a re-consideration due to the recent discovery of 5-hydroxymethylcytosine, a new DNA epigenetic mark present on genome. A recapitulation of the factors involved in the regulation of DNA methylation and the changes occurring in aging will be described in this review also considering the data available on 5 hmC., (Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.)
- Published
- 2015
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236. MARK-AGE biomarkers of ageing.
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Bürkle A, Moreno-Villanueva M, Bernhard J, Blasco M, Zondag G, Hoeijmakers JH, Toussaint O, Grubeck-Loebenstein B, Mocchegiani E, Collino S, Gonos ES, Sikora E, Gradinaru D, Dollé M, Salmon M, Kristensen P, Griffiths HR, Libert C, Grune T, Breusing N, Simm A, Franceschi C, Capri M, Talbot D, Caiafa P, Friguet B, Slagboom PE, Hervonen A, Hurme M, and Aspinall R
- Subjects
- European Union, Female, Humans, Male, Aging metabolism, Biomarkers metabolism
- Abstract
Many candidate biomarkers of human ageing have been proposed in the scientific literature but in all cases their variability in cross-sectional studies is considerable, and therefore no single measurement has proven to serve a useful marker to determine, on its own, biological age. A plausible reason for this is the intrinsic multi-causal and multi-system nature of the ageing process. The recently completed MARK-AGE study was a large-scale integrated project supported by the European Commission. The major aim of this project was to conduct a population study comprising about 3200 subjects in order to identify a set of biomarkers of ageing which, as a combination of parameters with appropriate weighting, would measure biological age better than any marker in isolation., (Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.)
- Published
- 2015
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237. 5mC-hydroxylase activity is influenced by the PARylation of TET1 enzyme.
- Author
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Ciccarone F, Valentini E, Zampieri M, and Caiafa P
- Subjects
- 5-Methylcytosine analogs & derivatives, Amino Acid Sequence, Catalytic Domain, Cytosine metabolism, DNA Damage, DNA Methylation, Enzyme-Linked Immunosorbent Assay, Epigenesis, Genetic, Glutathione Transferase metabolism, HEK293 Cells, Humans, Immunoprecipitation, Molecular Sequence Data, Poly (ADP-Ribose) Polymerase-1, Protein Processing, Post-Translational, Recombinant Proteins metabolism, Sequence Homology, Amino Acid, Cytosine analogs & derivatives, Gene Expression Regulation, Neoplastic, Mixed Function Oxygenases metabolism, Poly(ADP-ribose) Polymerases metabolism, Proto-Oncogene Proteins metabolism
- Abstract
5-hydroxymethylcytosine is a new epigenetic modification deriving from the oxidation of 5-methylcytosine by the TET hydroxylase enzymes. DNA hydroxymethylation drives DNA demethylation events and is involved in the control of gene expression. Deregulation of TET enzymes causes developmental defects and is associated with pathological conditions such as cancer. Little information thus far is available on the regulation of TET activity by post-translational modifications. Here we show that TET1 protein is able to interact with PARP-1/ARTD1 enzyme and is target of both noncovalent and covalent PARylation. In particular, we have demonstrated that the noncovalent binding of ADP-ribose polymers with TET1 catalytic domain decreases TET1 hydroxylase activity while the covalent PARylation stimulates TET1 enzyme. In addition, TET1 activates PARP-1/ARTD1 independently of DNA breaks. Collectively, our results highlight a complex interplay between PARylation and TET1 which may be helpful in coordinating the multiple biological roles played by 5-hydroxymethylcytosine and TET proteins.
- Published
- 2015
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238. PARP inhibitor ABT-888 affects response of MDA-MB-231 cells to doxorubicin treatment, targeting Snail expression.
- Author
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Mariano G, Ricciardi MR, Trisciuoglio D, Zampieri M, Ciccarone F, Guastafierro T, Calabrese R, Valentini E, Tafuri A, Del Bufalo D, Caiafa P, and Reale A
- Subjects
- Antibiotics, Antineoplastic pharmacology, Apoptosis drug effects, Apoptosis genetics, Blotting, Western, Breast Neoplasms genetics, Breast Neoplasms metabolism, Breast Neoplasms pathology, Cell Line, Tumor, Female, Gene Expression Regulation, Neoplastic drug effects, Humans, Inositol 1,4,5-Trisphosphate Receptors genetics, MCF-7 Cells, Poly (ADP-Ribose) Polymerase-1, Poly(ADP-ribose) Polymerases genetics, Promoter Regions, Genetic genetics, Protein Binding, Reverse Transcriptase Polymerase Chain Reaction, Snail Family Transcription Factors, Transcription Factors genetics, Benzimidazoles pharmacology, Doxorubicin pharmacology, Poly(ADP-ribose) Polymerase Inhibitors pharmacology, Poly(ADP-ribose) Polymerases metabolism, Transcription Factors metabolism
- Abstract
To overcome cancer cells resistance to pharmacological therapy, the development of new therapeutic approaches becomes urgent. For this purpose, the use of poly(ADP-ribose) polymerase (PARP) inhibitors in combination with other cytotoxic agents could represent an efficacious strategy. Poly(ADP-ribosyl)ation (PARylation) is a post-translational modification that plays a well characterized role in the cellular decisions of life and death. Recent findings indicate that PARP-1 may control the expression of Snail, the master gene of epithelial-mesenchymal transition (EMT). Snail is highly represented in different resistant tumors, functioning as a factor regulating anti-apoptotic programmes. MDA-MB-231 is a Snail-expressing metastatic breast cancer cell line, which exhibits chemoresistance properties when treated with damaging agents. In this study, we show that the PARP inhibitor ABT-888 was capable to modulate the MDA-MB-231 cell response to doxorubicin, leading to an increase in the rate of apoptosis. Our further results indicate that PARP-1 controlled Snail expression at transcriptional level in cells exposed to doxorubicin. Given the increasing interest in the employment of PARP inhibitors as chemotherapeutic adjuvants, our in vitro results suggest that one of the mechanisms through which PARP inhibition can chemosensitize cancer cells in vivo, is targeting Snail expression thus promoting apoptosis.
- Published
- 2015
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239. Poly(ADP-ribosyl)ation is involved in the epigenetic control of TET1 gene transcription.
- Author
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Ciccarone F, Valentini E, Bacalini MG, Zampieri M, Calabrese R, Guastafierro T, Mariano G, Reale A, Franceschi C, and Caiafa P
- Subjects
- Adenosine Diphosphate metabolism, DNA Methylation genetics, DNA-Binding Proteins genetics, Female, Gene Expression Regulation, Neoplastic, Histones genetics, Homeodomain Proteins genetics, Homeodomain Proteins metabolism, Humans, Jurkat Cells, MCF-7 Cells, Mixed Function Oxygenases, Poly(ADP-ribose) Polymerases genetics, Proto-Oncogene Proteins genetics, Transcription, Genetic genetics, Breast Neoplasms genetics, DNA-Binding Proteins metabolism, Epigenesis, Genetic, Poly(ADP-ribose) Polymerases metabolism, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma genetics, Proto-Oncogene Proteins metabolism
- Abstract
TET enzymes are the epigenetic factors involved in the formation of the sixth DNA base 5-hydroxymethylcytosine, whose deregulation has been associated with tumorigenesis. In particular, TET1 acts as tumor suppressor preventing cell proliferation and tumor metastasis and it has frequently been found down-regulated in cancer. Thus, considering the importance of a tight control of TET1 expression, the epigenetic mechanisms involved in the transcriptional regulation of TET1 gene are here investigated. The involvement of poly(ADP-ribosyl)ation in the control of DNA and histone methylation on TET1 gene was examined. PARP activity is able to positively regulate TET1 expression maintaining a permissive chromatin state characterized by DNA hypomethylation of TET1 CpG island as well as high levels of H3K4 trimethylation. These epigenetic modifications were affected by PAR depletion causing TET1 down-regulation and in turn reduced recruitment of TET1 protein on HOXA9 target gene. In conclusion, this work shows that PARP activity is a transcriptional regulator of TET1 gene through the control of epigenetic events and it suggests that deregulation of these mechanisms could account for TET1 repression in cancer.
- Published
- 2014
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240. The epigenetic factor BORIS/CTCFL regulates the NOTCH3 gene expression in cancer cells.
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Zampieri M, Ciccarone F, Palermo R, Cialfi S, Passananti C, Chiaretti S, Nocchia D, Talora C, Screpanti I, and Caiafa P
- Subjects
- Cells, Cultured, DNA Methylation, Humans, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma genetics, Promoter Regions, Genetic, Receptor, Notch3, DNA-Binding Proteins physiology, Epigenesis, Genetic, Gene Expression Regulation, Neoplastic, Receptors, Notch genetics
- Abstract
Aberrant upregulation of NOTCH3 gene plays a critical role in cancer pathogenesis. However, the underlying mechanisms are still unknown. We tested here the hypothesis that aberrant epigenetic modifications in the NOTCH3 promoter region might account for its upregulation in cancer cells. We compared DNA and histone methylation status of NOTCH3 promoter region in human normal blood cells and T cell acute lymphoblastic leukemia (T-ALL) cell lines, differentially expressing NOTCH3. We found that histone methylation, rather than DNA hypomethylation, contributes towards establishing an active chromatin status of NOTCH3 promoter in NOTCH3 overexpressing cancer cells. We discovered that the chromatin regulator protein BORIS/CTCFL plays an important role in regulating NOTCH3 gene expression. We observed that BORIS is present in T-ALL cell lines as well as in cell lines derived from several solid tumors overexpressing NOTCH3. Moreover, BORIS targets NOTCH3 promoter in cancer cells and it is able to induce and to maintain a permissive/active chromatin conformation. Importantly, the association between NOTCH3 overexpression and BORIS presence was confirmed in primary T-ALL samples from patients at the onset of the disease. Overall, our results provide novel insights into the determinants of NOTCH3 overexpression in cancer cells, by revealing a key role for BORIS as the main mediator of transcriptional deregulation of NOTCH3., (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Published
- 2014
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241. Pharmacological inhibition of poly(ADP-ribose) polymerase-1 modulates resistance of human glioblastoma stem cells to temozolomide.
- Author
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Tentori L, Ricci-Vitiani L, Muzi A, Ciccarone F, Pelacchi F, Calabrese R, Runci D, Pallini R, Caiafa P, and Graziani G
- Subjects
- Cell Line, Tumor, CpG Islands, DNA Methylation, Dacarbazine pharmacology, Glioblastoma genetics, Glioblastoma mortality, Humans, O(6)-Methylguanine-DNA Methyltransferase genetics, O(6)-Methylguanine-DNA Methyltransferase metabolism, Poly (ADP-Ribose) Polymerase-1, Poly(ADP-ribose) Polymerases metabolism, Promoter Regions, Genetic, Temozolomide, Antineoplastic Agents, Alkylating pharmacology, Dacarbazine analogs & derivatives, Drug Resistance, Neoplasm, Glioblastoma metabolism, Neoplastic Stem Cells drug effects, Neoplastic Stem Cells metabolism, Poly(ADP-ribose) Polymerase Inhibitors
- Abstract
Background: Chemoresistance of glioblastoma multiforme (GBM) has been attributed to the presence within the tumor of cancer stem cells (GSCs). The standard therapy for GBM consists of surgery followed by radiotherapy and the chemotherapeutic agent temozolomide (TMZ). However, TMZ efficacy is limited by O6-methylguanine-DNA-methyltransferase (MGMT) and Mismatch Repair (MMR) functions. Strategies to counteract TMZ resistance include its combination with poly(ADP-ribose) polymerase inhibitors (PARPi), which hamper the repair of N-methylpurines. PARPi are also investigated as monotherapy for tumors with deficiency of homologous recombination (HR). We have investigated whether PARPi may restore GSC sensitivity to TMZ or may be effective as monotherapy., Methods: Ten human GSC lines were assayed for MMR proteins, MGMT and PARP-1 expression/activity, MGMT promoter methylation and sensitivity to TMZ or PARPi, alone and in combination. Since PTEN defects are frequently detected in GBM and may cause HR dysfunction, PTEN expression was also analyzed. The statistical analysis of the differences in drug sensitivity among the cell lines was performed using the ANOVA and Bonferroni's post-test or the non-parametric Kruskal-Wallis analysis and Dunn's post-test for multiple comparisons. Synergism between TMZ and PARPi was analyzed by the median-effect method of Chou and Talalay. Correlation analyses were done using the Spearman's rank test., Results: All GSCs were MMR-proficient and resistance to TMZ was mainly associated with high MGMT activity or low proliferation rate. MGMT promoter hypermethylation of GSCs correlated both with low MGMT activity/expression (Spearman's test, P = 0.004 and P = 0.01) and with longer overall survival of GBM patients (P = 0.02). Sensitivity of each GSC line to PARPi as single agent did not correlate with PARP-1 or PTEN expression. Notably, PARPi and TMZ combination exerted synergistic antitumor effects in eight out of ten GSC lines and the TMZ dose reduction achieved significantly correlated with the sensitivity of each cell line to PARPi as single agent (P = 0.01)., Conclusions: The combination of TMZ with PARPi may represent a valuable strategy to reverse GSC chemoresistance.
- Published
- 2014
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242. Essential role of poly(ADP-ribosyl)ation in cocaine action.
- Author
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Scobie KN, Damez-Werno D, Sun H, Shao N, Gancarz A, Panganiban CH, Dias C, Koo J, Caiafa P, Kaufman L, Neve RL, Dietz DM, Shen L, and Nestler EJ
- Subjects
- Animals, Behavior, Animal drug effects, Chromatin Immunoprecipitation, Cocaine administration & dosage, Dendritic Spines drug effects, Dendritic Spines metabolism, Gene Expression Regulation, Enzymologic drug effects, Genome genetics, Immunoglobulin G metabolism, Male, Membrane Proteins metabolism, Mice, Mice, Inbred C57BL, Nucleus Accumbens drug effects, Nucleus Accumbens enzymology, Poly(ADP-ribose) Polymerases genetics, Poly(ADP-ribose) Polymerases metabolism, Protein Binding drug effects, RNA, Messenger genetics, RNA, Messenger metabolism, Rats, Substrate Specificity drug effects, Transcription, Genetic drug effects, Cocaine pharmacology, Poly Adenosine Diphosphate Ribose metabolism
- Abstract
Many of the long-term effects of cocaine on the brain's reward circuitry have been shown to be mediated by alterations in gene expression. Several chromatin modifications, including histone acetylation and methylation, have been implicated in this regulation, but the effect of other histone modifications remains poorly understood. Poly(ADP-ribose) polymerase-1 (PARP-1), a ubiquitous and abundant nuclear protein, catalyzes the synthesis of a negatively charged polymer called poly(ADP-ribose) or PAR on histones and other substrate proteins and forms transcriptional regulatory complexes with several other chromatin proteins. Here, we identify an essential role for PARP-1 in cocaine-induced molecular, neural, and behavioral plasticity. Repeated cocaine administration, including self-administration, increased global levels of PARP-1 and its mark PAR in mouse nucleus accumbens (NAc), a key brain reward region. Using PARP-1 inhibitors and viral-mediated gene transfer, we established that PARP-1 induction in NAc mediates enhanced behavioral responses to cocaine, including increased self-administration of the drug. Using chromatin immunoprecipitation sequencing, we demonstrated a global, genome-wide enrichment of PARP-1 in NAc of cocaine-exposed mice and identified several PARP-1 target genes that could contribute to the lasting effects of cocaine. Specifically, we identified sidekick-1--important for synaptic connections during development--as a critical PARP-1 target gene involved in cocaine's behavioral effects as well as in its ability to induce dendritic spines on NAc neurons. These findings establish the involvement of PARP-1 and PARylation in the long-term actions of cocaine.
- Published
- 2014
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243. ADP-ribose polymer depletion leads to nuclear Ctcf re-localization and chromatin rearrangement(1).
- Author
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Guastafierro T, Catizone A, Calabrese R, Zampieri M, Martella O, Bacalini MG, Reale A, Di Girolamo M, Miccheli M, Farrar D, Klenova E, Ciccarone F, and Caiafa P
- Subjects
- Active Transport, Cell Nucleus, Amino Acid Substitution, Animals, CCCTC-Binding Factor, Cell Line, Cell Nucleus metabolism, Chromatin Assembly and Disassembly, DNA Methylation, Gene Knockdown Techniques, Glycoside Hydrolases genetics, Glycoside Hydrolases metabolism, Lamins metabolism, Mice, Mutagenesis, Site-Directed, Mutant Proteins genetics, Mutant Proteins metabolism, Poly (ADP-Ribose) Polymerase-1, Poly(ADP-ribose) Polymerase Inhibitors, Poly(ADP-ribose) Polymerases genetics, Poly(ADP-ribose) Polymerases metabolism, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Repressor Proteins antagonists & inhibitors, Repressor Proteins genetics, Adenosine Diphosphate Ribose metabolism, Repressor Proteins metabolism
- Abstract
Ctcf (CCCTC-binding factor) directly induces Parp [poly(ADP-ribose) polymerase] 1 activity and its PARylation [poly(ADPribosyl)ation] in the absence of DNA damage. Ctcf, in turn, is a substrate for this post-synthetic modification and as such it is covalently and non-covalently modified by PARs (ADP-ribose polymers). Moreover, PARylation is able to protect certain DNA regions bound by Ctcf from DNA methylation. We recently reported that de novo methylation of Ctcf target sequences due to overexpression of Parg [poly(ADP-ribose)glycohydrolase] induces loss of Ctcf binding. Considering this, we investigate to what extent PARP activity is able to affect nuclear distribution of Ctcf in the present study. Notably, Ctcf lost its diffuse nuclear localization following PAR (ADP-ribose polymer) depletion and accumulated at the periphery of the nucleus where it was linked with nuclear pore complex proteins remaining external to the perinuclear Lamin B1 ring. We demonstrated that PAR depletion-dependent perinuclear localization of Ctcf was due to its blockage from entering the nucleus. Besides Ctcf nuclear delocalization, the outcome of PAR depletion led to changes in chromatin architecture. Immunofluorescence analyses indicated DNA redistribution, a generalized genomic hypermethylation and an increase of inactive compared with active chromatin marks in Parg-overexpressing or Ctcf-silenced cells. Together these results underline the importance of the cross-talk between Parp1 and Ctcf in the maintenance of nuclear organization.
- Published
- 2013
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244. Why is DNA methylation of Igf2 CpG island shore affected during ageing?
- Author
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Caiafa P and Zampieri M
- Subjects
- Female, Humans, Male, Aging metabolism, DNA Methylation, Insulin-Like Growth Factor II genetics, RNA, Long Noncoding genetics
- Published
- 2012
- Full Text
- View/download PDF
245. Methylation-dependent PAD2 upregulation in multiple sclerosis peripheral blood.
- Author
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Calabrese R, Zampieri M, Mechelli R, Annibali V, Guastafierro T, Ciccarone F, Coarelli G, Umeton R, Salvetti M, and Caiafa P
- Subjects
- Adult, Brain enzymology, Brain metabolism, CpG Islands genetics, DNA Methylation, Female, Humans, Hydrolases blood, Hydrolases metabolism, Leukocytes, Mononuclear metabolism, Male, Middle Aged, Multiple Sclerosis enzymology, Myelin Basic Protein genetics, Myelin Basic Protein metabolism, Protein-Arginine Deiminase Type 2, Protein-Arginine Deiminases, Up-Regulation, Hydrolases genetics, Leukocytes, Mononuclear enzymology, Multiple Sclerosis genetics
- Abstract
Background: Peptidylarginine deiminase 2 (PAD2) and peptidylarginine deiminase 4 (PAD4) are two members of PAD family which are over-expressed in the multiple sclerosis (MS) brain. Through its enzymatic activity PAD2 converts myelin basic protein (MBP) arginines into citrullines - an event that may favour autoimmunity - while peptidylarginine deiminase 4 (PAD4) is involved in chromatin remodelling., Objectives: Our aim was to verify whether an altered epigenetic control of PAD2, as already shown in the MS brain, can be observed in peripheral blood mononuclear cells (PBMCs) of patients with MS since some of these cells also synthesize MBP., Methods: The expression of most suitable reference genes and of PAD2 and PAD4 was assessed by qPCR. Analysis of DNA methylation was performed by bisulfite method., Results: The comparison of PAD2 expression level in PBMCs from patients with MS vs. healthy donors showed that, as well as in the white matter of MS patients, the enzyme is significantly upregulated in affected subjects. Methylation pattern analysis of a CpG island located in the PAD2 promoter showed that over-expression is associated with promoter demethylation., Conclusion: Defective regulation of PAD2 in the periphery, without the immunological shelter of the blood-brain barrier, may contribute to the development of the autoimmune responses in MS.
- Published
- 2012
- Full Text
- View/download PDF
246. A subset of chronic lymphocytic leukemia patients display reduced levels of PARP1 expression coupled with a defective irradiation-induced apoptosis.
- Author
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Bacalini MG, Tavolaro S, Peragine N, Marinelli M, Santangelo S, Del Giudice I, Mauro FR, Di Maio V, Ricciardi MR, Caiafa P, Chiaretti S, Foà R, Guarini A, and Reale A
- Subjects
- Apoptosis Regulatory Proteins genetics, Apoptosis Regulatory Proteins metabolism, DNA Damage, DNA, Neoplasm radiation effects, Gamma Rays, Genes, p53, Humans, Immunoglobulin Heavy Chains genetics, Immunoglobulin Variable Region genetics, Leukemia, Lymphocytic, Chronic, B-Cell genetics, Leukemia, Lymphocytic, Chronic, B-Cell metabolism, Mutation, Neoplasm Proteins genetics, Neoplasm Proteins physiology, Oligonucleotide Array Sequence Analysis, Poly (ADP-Ribose) Polymerase-1, Poly(ADP-ribose) Polymerases genetics, Poly(ADP-ribose) Polymerases physiology, RNA, Messenger biosynthesis, RNA, Neoplasm biosynthesis, Repressor Proteins genetics, Repressor Proteins metabolism, Treatment Outcome, Tumor Cells, Cultured metabolism, Tumor Cells, Cultured radiation effects, Tumor Suppressor Protein p53 biosynthesis, Apoptosis radiation effects, Gene Expression Regulation, Leukemic radiation effects, Leukemia, Lymphocytic, Chronic, B-Cell pathology, Neoplasm Proteins biosynthesis, Poly(ADP-ribose) Polymerases biosynthesis
- Abstract
Chronic lymphocytic leukemia (CLL) is a heterogeneous disease characterized by defects in the DNA damage response and apoptosis. Among the factors involved in these pathways, we focused on the enzyme poly(ADP-ribose) polymerase 1 (PARP1) and on its substrate Che-1 by evaluating their basal expression and functional changes upon irradiation (IR). Microarray experiments were performed on 98 untreated CLL cases. Next, freshly isolated primary cells from 21 untreated patients were analyzed for in vitro response to irradiation through Western blot, PARP activity assay, Annexin-V analysis, and PARP1 basal expression by quantitative polymerase chain reaction. Microarray analysis showed that PARP1 and CHE1 were constitutively expressed in CLL and had a high degree of correlation with each other and with TP53. PARP1 and TP53 downmodulation was associated with worse clinical outcomes, especially in TP53-mutated cases. Next, CLL samples from 21 untreated patients were classified as responders and nonresponders based on IR-induced PARP1 cleavage. Notably, while responder samples were characterized by Che-1 and p53 induction at 8 hours and reduction at 24 hours post-IR, nonresponders included both samples with p53 dysfunctions and cases with a normal IR-induced Che-1 and/or p53 response. Finally, we observed that PARP1 was downregulated in nonresponder vs responder samples and that its basal expression was positively correlated with PARP1 cleavage after IR. In conclusion, we showed that reduced expression of PARP1 is associated with an impairment of CLL responsiveness to cell death., (Copyright © 2012 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
247. ADP-ribose polymers localized on Ctcf-Parp1-Dnmt1 complex prevent methylation of Ctcf target sites.
- Author
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Zampieri M, Guastafierro T, Calabrese R, Ciccarone F, Bacalini MG, Reale A, Perilli M, Passananti C, and Caiafa P
- Subjects
- CCCTC-Binding Factor, CpG Islands physiology, DNA (Cytosine-5-)-Methyltransferase 1, DNA Methylation, Epigenesis, Genetic, Multiprotein Complexes metabolism, DNA (Cytosine-5-)-Methyltransferases metabolism, Poly Adenosine Diphosphate Ribose metabolism, Poly(ADP-ribose) Polymerases metabolism, Repressor Proteins metabolism
- Abstract
PARylation [poly(ADP-ribosyl)ation] is involved in the maintenance of genomic methylation patterns through its control of Dnmt1 [DNA (cytosine-5)-methyltransferase 1] activity. Our previous findings indicated that Ctcf (CCCTC-binding factor) may be an important player in key events whereby PARylation controls the unmethylated status of some CpG-rich regions. Ctcf is able to activate Parp1 [poly(ADP-ribose) polymerase 1], which ADP-ribosylates itself and, in turn, inhibits DNA methylation via non-covalent interaction between its ADP-ribose polymers and Dnmt1. By such a mechanism, Ctcf may preserve the epigenetic pattern at promoters of important housekeeping genes. The results of the present study showed Dnmt1 as a new protein partner of Ctcf. Moreover, we show that Ctcf forms a complex with Dnmt1 and PARylated Parp1 at specific Ctcf target sequences and that PARylation is responsible for the maintenance of the unmethylated status of some Ctcf-bound CpGs. We suggest a mechanism by which Parp1, tethered and activated at specific DNA target sites by Ctcf, preserves their methylation-free status.
- Published
- 2012
- Full Text
- View/download PDF
248. Poly(ADP-ribosyl)ation acts in the DNA demethylation of mouse primordial germ cells also with DNA damage-independent roles.
- Author
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Ciccarone F, Klinger FG, Catizone A, Calabrese R, Zampieri M, Bacalini MG, De Felici M, and Caiafa P
- Subjects
- Analysis of Variance, Animals, Blotting, Western, DNA (Cytosine-5-)-Methyltransferase 1, DNA (Cytosine-5-)-Methyltransferases genetics, DNA (Cytosine-5-)-Methyltransferases metabolism, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Embryo, Mammalian cytology, Embryo, Mammalian embryology, Female, Gene Expression Regulation, Developmental, Male, Mice, Poly (ADP-Ribose) Polymerase-1, Poly Adenosine Diphosphate Ribose metabolism, Poly(ADP-ribose) Polymerases genetics, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Time Factors, DNA Damage, DNA Methylation, Embryo, Mammalian metabolism, Germ Cells metabolism, Poly(ADP-ribose) Polymerases metabolism
- Abstract
Poly(ADP-ribosyl)ation regulates chromatin structure and transcription driving epigenetic events. In particular, Parp1 is able to directly influence DNA methylation patterns controlling transcription and activity of Dnmt1. Here, we show that ADP-ribose polymer levels and Parp1 expression are noticeably high in mouse primordial germ cells (PGCs) when the bulk of DNA demethylation occurs during germline epigenetic reprogramming in the embryo. Notably, Parp1 activity is stimulated in PGCs even before its participation in the DNA damage response associated with active DNA demethylation. We demonstrate that PARP inhibition impairs both genome-wide and locus-specific DNA methylation erasure in PGCs. Moreover, we evidence that impairment of PARP activity causes a significant reduction of expression of the gene coding for Tet1 hydroxylases involved in active DNA demethylation. Taken together these results demonstrate new and adjuvant roles of poly(ADP-ribosyl)ation during germline DNA demethylation and suggest its possible more general involvement in genome reprogramming.
- Published
- 2012
- Full Text
- View/download PDF
249. Proteome analysis of protein partners to nucleosomes containing canonical H2A or the variant histones H2A.Z or H2A.X.
- Author
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Fujimoto S, Seebart C, Guastafierro T, Prenni J, Caiafa P, and Zlatanova J
- Subjects
- Genetic Variation genetics, HeLa Cells, Histones chemistry, Histones genetics, Histones isolation & purification, Humans, Proteome metabolism, Histones metabolism, Nuclear Proteins analysis, Nuclear Proteins metabolism, Nucleosomes chemistry, Nucleosomes metabolism, Proteome analysis
- Abstract
Although the existence of histone variants has been known for quite some time, only recently are we grasping the breadth and diversity of the cellular processes in which they are involved. Of particular interest are the two variants of histone H2A, H2A.Z and H2A.X because of their roles in regulation of gene expression and in DNA double-strand break repair, respectively. We hypothesize that nucleosomes containing these variants may perform their distinct functions by interacting with different sets of proteins. Here, we present our proteome analysis aimed at identifying protein partners that interact with nucleosomes containing H2A.Z, H2A.X or their canonical H2A counterpart. Our development of a nucleosome-pull down assay and analysis of the recovered nucleosome-interacting proteins by mass spectrometry allowed us to directly compare nuclear partners of these variant-containing nucleosomes to those containing canonical H2A. To our knowledge, our data represent the first systematic analysis of the H2A.Z and H2A.X interactome in the context of nucleosome structure.
- Published
- 2012
- Full Text
- View/download PDF
250. Poly(ADP-ribosyl)ation affects stabilization of Che-1 protein in response to DNA damage.
- Author
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Bacalini MG, Di Lonardo D, Catizone A, Ciccarone F, Bruno T, Zampieri M, Guastafierro T, Calabrese R, Fanciulli M, Passananti C, Caiafa P, and Reale A
- Subjects
- Animals, Antineoplastic Agents pharmacology, Apoptosis Regulatory Proteins genetics, Ataxia Telangiectasia Mutated Proteins, Cell Cycle Proteins metabolism, Cell Line, DNA-Binding Proteins metabolism, Enzyme Activation drug effects, Gene Expression Regulation, Neoplastic drug effects, Humans, Mice, Promoter Regions, Genetic, Protein Binding, Protein Serine-Threonine Kinases metabolism, Protein Stability, Proto-Oncogene Proteins p21(ras) genetics, Recombinant Fusion Proteins genetics, Tumor Suppressor Proteins metabolism, Apoptosis Regulatory Proteins metabolism, DNA Damage drug effects, Poly(ADP-ribose) Polymerases metabolism, Recombinant Fusion Proteins metabolism
- Abstract
Poly(ADP-ribose) polymerase 1 (PARP-1) catalyzes a post-translational modification that plays a crucial role in coordinating the signalling cascade in response to stress stimuli. During the DNA damage response, phosphorylation by ataxia telangiectasia mutated (ATM) kinase and checkpoint kinase Chk2 induces the stabilization of Che-1 protein, which is critical for the maintenance of G2/M arrest. In this study we showed that poly(ADP-ribosyl)ation, beyond phosphorylation, is involved in the regulation of Che-1 stabilization following DNA damage. We demonstrated that Che-1 accumulation upon doxorubicin treatment is reduced after the inhibition of PARP activity in HCT116 cells and in PARP-1 knock-out or silenced cells. In accordance, impairment in Che-1 accumulation by PARP inhibition reduced Che-1 occupancy at p21 promoter and affected the expression of the corresponding gene. Epistasis experiments showed that the effect of poly(ADP-ribosyl)ation on Che-1 stabilization is independent from ATM kinase activity. Indeed we demonstrated that Che-1 protein co-immunoprecipitates with ADP-ribose polymers and that PARP-1 directly interacts with Che-1, promoting its modification in vitro and in vivo., (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
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