0000000000016552
AUTHOR
Anna Maria Rinaldi
RNA binding proteins: putative regulative role in mitochondrial proteins expression.
Mitochondrial protein expression in Ratt and in human cells
In the present paper, we analyzed some aspects of the post-transcriptional regulation of two COX (cy-tochrome c oxidase) subunits, i.e. mitochondrion-encoded COXIII and nucleus-encoded COXIV. In particular, by T1 RNase protection assays, we found two proteins, present in mitochondrial extracts from adult rat brain and testis, able to bind COXIII mRNA. We also found cytoplasmic proteins present in testis, kidney and heart extracts that bind COXIV mRNA. Moreover, to study the expression of mitochondrial proteins in tumor cells, we quantitated COXIV and Hsp60 chaperonin abundance in two epithelial cell lines from human breast, one neoplastic (MDA-MB231) and the other immortalized and non-tumor…
Mitochondrial proteins regulation in Rattus norvegicus and human cells
Modificazioni nelle comunità fitobentoniche di fouling in relazione all’impatto antropico ed alla geometria del substrato
Questo studio è stato condotto nella primavera del 2006 in due località del Golfo di Palermo (Arenella e Bandita) a differente grado di impatto antropico. All’interno di ogni località, sono stati scelti siti a differente inclinazione (orizzontale vs verticale). Sono stati campionati i popolamenti mesolitorali dei massi frangiflutti che, insieme a quelli della frangia infralitorale, risentono maggiormente dell’eventuale presenza di inquinamento organico. Le comunità algali nelle due località hanno mostrato differenze in funzione delle diverse caratteristiche ambientali e dei diversi livelli di impatto antropico, in termini di copertura %, strutturazione e diversità specifica. Sono stati inol…
Effects of cadmium chloride on some mitochondria-related activity and gene expression of human MDA-MB231 breast tumor cells.
It was reported that cadmium is able to exert a cytotoxic effect on tumor MDA-MB231 cells, which shows signs of "non-classical" apoptosis and is characterized by drastic changes in gene expression pattern. In this study, we have extended our knowledge of metal-breast cancer cell interactions by analyzing some mitochondria-related aspects of the stress response to CdCl(2) at either 5 or 50 microM 24- or 96-h exposure, by cytochemical, conventional PCR and Northern/Western blot techniques. We demonstrated that (i) no modification of the mitochondrial mass was detectable due to CdCl(2) exposure; (ii) the respiration activity appeared to be increased after 96-h exposures, while the production o…
Male infertility and mitochondrial DNA
The mitochondrial machinery plays a key role in the energy production and maintenance of spermatozoa motility. In this paper 200 idiopathic oligo-asthenozoospermic patients were classified on the basis of rapid progressive motility ("a") and sperm concentration. Mitochondrial enzymatic activity was studied and correlated to the viability of sperm cells. Mitochondrial DNA purified from both motile and non-motile sperm of the same individuals was amplificated using PCR. Results suggested that only motile sperm have organelles functional in oxygen consumption, unequivocally demonstrating that motility depends on the mitochondrial activity. Mitochondrial DNA of oligo-asthenozoospermic patients …
Sea Urchin Mitochondrial Matrix Contains a 56-kDa Chaperonine-like Protein
Abstract Paracentrotus lividus mitochondrial matrix contains a constitutive hsp of 56-KDa which cross reacts with a serum anti-hsp-60 chaperonine from yeast mitochondria. The localization of hsps preexisting or newly synthesized in different subcellular fractions of gastrula embryos is also analyzed by two-dimensional electrophoresis.
Mitochondrial compartment: a possible target of cadmium effects on breast epithelial cells.
Cadmium–breast epithelial cell interactions were studied by analyzing some mitochondria-related aspects of stress response. We treated immortalized non-tumor breast cells with 5 or 50 μM CdCl2 for 24 or 96 h demonstrating that the exposure did not cause a significant mitochondrial proliferation, while it induced a significant increase in the respiratory activity and mitochondrial polarization. In addition, we found that hsp60 was up-regulated while hsp70 and COXII and COXIV were down-regulated. The mRNA for hsp70 remained constant and only the inducible form of the 70-kDa heat shock protein was over expressed. The mRNAs for COXII and COXIV remained constant after 24 h and increased after lo…
Possible mechanisms regulating the expression of nuclear and mitochondrial genes
Cytochemical and molecular analyses on mitochondria of immortalized and neoplastic epithelial cells of the human breast after cadmium treatments
Cadmium induces an apoptotic response in sea urchin embryos.
Cadmium is a heavy metal toxic for living organisms even at low concentrations. It does not have any biological role, and since it is a permanent metal ion, it is accumulated by many organisms. In the present paper we have studied the apoptotic effects of continuous exposure to subacute/sublethal cadmium concentrations on a model system: Paracentrotus lividus embryos. We demonstrated, by atomic absorption spectrometry, that the intracellular amount of metal increased during exposure time. We found, using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay, that long treatments with cadmium triggered a severe DNA fragmentation. We demonstrated, by immunocytochemistry …
In vitro incorporation of amino acids into proteins stimulated by RNA from unfertilized sea urchin eggs.
Possible mechanisms regulating the expression of nuclear and mitochondrial genes
Apoptotic pathway induced by cadmium in sea urchin embryos
Nuclear-mitochondrial interaction.
The biogenesis of mitochondria depends on the coordinated expression of nuclear and mitochondrial genomes. Consequently, the control of mitochondrial biogenesis and function depends on extremely complex processes requiring a variety of well orchestrated regulatory mechanisms. It is clear that the interplay of transcription factors and coactivators contributes to the expression of both nuclear and mitochondrial respiratory genes. In addition, the regulation of mitochondria biogenesis depends on proteins that, interacting with messenger RNAs for mitochondrial proteins, influence their metabolism and expression. Moreover, a tight regulation of the import and final assembly of mitochondrial pro…
MITOCHONDRIA DURING SEA URCHIN OVOGENESIS
Effetti dell’impatto antropico e della geometria del substrato su struttura e distribuzione delle comunità bentoniche di fouling
Effect of cadmium chloride on some mitochondria-related activity and gene expression of human MDA-MB231 breast tumor cells
Regolazione dell’espressione delle subunità III e IV della citocromo c ossidasi
Cadmium and mitochondria
The heavy metal cadmium (Cd) a pollutant associated with several modern industrial processes, is absorbed in significant quantities from cigarette smoke, water, food and air contaminations. It is known to have numerous undesirable effects on health in both experimental animals and humans, targeting kidney, liver and vascular system. The molecular mechanism accounting for most of the biological effects of Cd are not well-understood and the toxicity targets are largely unidentified. The present review focuses on important recent advances about the effects of cadmium on mitochondria of mammalian cells. Mitochondria are the proverbial powerhouses of the cell, running the fundamental biochemical…
A method for measuring mitochondrial mass and activity
Mitochondria, responsible for the energy-generating process essential for the cell metabolism, differ for the number, localization and activity in animal cells and tissues in relation to the energetic needs. Using fluorescent probes specific for mitochondria, Mitotracker Green (MTG) and Orange (MTO), and Confocal Laser-Scanning Microscope (CLSM), we elaborated a method to measure in vivo the mitochondrial mass and activity, in sea urchin Paracentrotus lividus eggs and embryos. The analysis of captured images, revealed a variation of mitochondrial distribution and an increase of activity after fertilization.
Study on endophytic strains of genus Acremonium in symptomless grapevines
Induction of apoptosis by cadmium in Paracentrotus lividus embryos and larvae
Induction of apoptosis by cadmium in sea urchin embryos: possible involvement of extrinsic pathway
Proteine di fusione, metodo di preparazione, e relativi anticorpi per la diagnosi di patologie mitocondriali
Proteins participating to the post-transcriptional regulation of the mitochondrial cytochrome c oxidase subunit IV via elements located in the 3′UTR
Abstract In developing rat brain cytochrome c oxidase subunit IV (COXIV) expression is also regulated at post-transcriptional level and two 3′UTR-COXIV RNA-binding factors have been identified. Here, we report the enrichment and identification of the factors from just born rat brains by affinity chromatography of biotinylated 3′UTR-COXIV RNA–protein complexes on streptavidin-conjugated paramagnetic particles. We successfully isolated two main proteins of about 86 and 42 kDa, whose sequences were highly attributable to Hsp90 and Actin. The purified proteins maintain RNA-binding ability and specificity for COXIV messenger and, interacting with the 3′UTR, then could negatively modulate mRNA tr…
Cytochemical and molecular analyses on mitochondria of immortalized and neoplastic epithelial cells from the human breast
STUDIO DELL’ESPRESSIONE DI GENI CODIFICANTI PROTEINE MITOCONDRIALI IN RATTUS NORVEGICUS.
Hsp 56 mRNA in Paracentrotus lividus embryos binds to a mitochondrial protein
We previously demonstrated that Paracentrotus lividus Hsp56 mitochondrial chaperonin is constitutively expressed during development, that it has a specific territorial distribution, both in normal and heat-shocked embryos, and that its amount increases after heat shock [Roccheri MC, Patti M, Agnello M, Gianguzza F, Carra E, Rinaldi AM. Localization of mitochondrial Hsp56 chaperonin during sea urchin development. Biochem Biophys Res Commun 2001;287:1093-98] and cadmium treatment [Roccheri MC, Agnello M, Boneventura R, Matranga V. Cadmium induces the expression of specific stress proteins in sea urchin embryos. Biochem Biophys Res Commun 2004;321:80-7]. In this study, we looked at Hsp56 mRNA …
Mitochondria and Development
Mitochondrial DNA deletion and male infertility
Abstract The term “male infertility” does not constitute a defined clinical syndrome, but rather, a collection of different conditions exhibiting a variety of aetiologies and varying diagnosis. In men, oligozoospermia, asthenozoospermia, teratozoospermia and azoospermia are the main causes of infertility. The present research is aimed to investigate if mtDNA deletions can cause sperm defects in idiopathic astenozoospermic patients with different sperm motility and sperm concentration. The incidence of deletions in mtDNA (mtDNA) was co-related with diagnostic categorization of male infertility when 6.6kb out of the 16.6kb of the mitochondrial genome was amplified from whole sperm samples, f…
STUDY ON AN ENDOPHYTIC POPULATION OF ACREMONIUM SP. IN SYMPTOMLESS GRAPEVINE
Hsp56 expression in Paracentrotus lividus embryos
Effetto del cadmio sull'attività mitocondriale e sull'espressione genica in cellule tumorali e immortalizzatedi epitelio ghiandolare mammario
Isolation and characterization of a Paracentrotus lividus cDNA encoding a stress-inducible chaperonin
Chaperonins are ubiquitous proteins that facilitate protein folding in an adenosine triphosphate–dependent manner. Here we report the isolation of a sea urchin cDNA (Plhsp60) coding for mitochondrial chaperonin (Cpn60), whose basal expression is further enhanced by heat shock. The described cDNA corresponds to a full-length mRNA encoding a protein of 582 amino acids, the first 32 of which constitute a putative mitochondrial targeting leader sequence. Comparative analysis has demonstrated that this protein is highly conserved in evolution.
Analysis of cytochrome C oxidase subunits III and IV expression in developing rat brain
Abstract Cytochrome c oxidase (COX) complex is built up with both nucleus- and mitochondrion-encoded subunits. Biogenesis and assembly of the complex thus requires fine cross-talk between the two compartments. In order to shed light on the regulation of nuclear–mitochondrial interactions, we studied the expression of COXIII (mitochondrion-encoded) and COXIV (nucleus-encoded) in adult rat tissues and rat developing brain. We found that the levels of COXIV protein and mRNA are not linearly related, thus suggesting a post-transcriptional mode of regulation. In agreement with this observation, we report the presence of a protein that specifically binds to the 3′-untranslated region of COXIV mRN…
Astenozoospermy and mitochondrial DNA
Male infertility, inability to fecundate the oocyte, is due to alterations of sperm characteristics (low number, low motility, altered morphology), to the quantity of semen or to the presence of alterations in the male genital line. 75% of infertile men deal with untreatable sub-fertility. Semen of 200 idiopathic astenozoospermic patients and individuals with normal seminal parameters as control, were analyzed. Results showed that mtDNA and Y chromosome regions from the motile fractions of patients and controls gave amplifications, while, the non-motile fractions of the same seminal samples did not give amplifications of ND5, ND6 and regions inside the D-loop (Carra E. et al.2002, 2004). We…
Localization of mitochondrial Hsp56 chaperonin during sea urchin development.
We have previously demonstrated that Paracentrotus lividus nuclear genome encodes for the heat shock inducible chaperonin homolog Hsp 56 (1) and that the mature protein is localized in the mitochondrial matrix (2). In this paper we report that constitutive Hsp56 is maternally inherited, in fact it is present in the in unfertilized eggs, and that it has a perinuclear specific localization during cleavage. In the later stages both the constitutive and the heat shock inducible chaperonin has a specific territorial distribution. Moreover following heat shock, the Hsp56 appears in the cytoplasm and in the postmitochondrial supernatant beside the mitochondrial fraction.
Analysis of cytochrome c oxidase subunits III and IV in developing rat brain
Analysis of cytochrome oxidase subunits III and IV expression in developing rat brain.
Mitochondria during sea urchin oogenesis.
SummarySea urchin represents an ideal model for studies on fertilization and early development, but the achievement of egg competence and mitochondrial behaviour during oogenesis remain to be enlightened. Oocytes of echinoid, such as sea urchin, unlike other echinoderms and other systems, complete meiotic maturation before fertilization. Mitochondria, the powerhouse of eukaryotic cells, contain a multi-copy of the maternally inherited genome, and are involved directly at several levels in the reproductive processes, as their functional status influences the quality of oocytes and contributes to fertilization and embryogenesis. In the present paper, we report our latest data on mitochondrial…
Centrifugation does not alter spatial distribution of `BEP4' mRNA in paracentrotus lividus EGG
AbstractParacentrotus lividus unfertilized eggs were centrifuged in a sucrose gradient, so to split each into two parts: a nucleated light fragment and an anucleated heavy fragment. Northern blot analyses utilizing a bep4 probe as animal marker and H2A histone gene and 12S-mit RNA as controls indicate that the eggs are elongated along the animal-vegetal axis during centrifugation and thereafter split into an animal and a vegetal half. Treatment of the eggs with colchicine before centrifugation abolishes the animal localization of bep4 mRNA.
Effetti del cadmio sugli embrioni di riccio di mare
Analysis of cytochrome c oxidase subunits III and IV expression in developing rat brain
Espressione del messaggero per l’Hsp56 in embrioni di Paracentrotus lividus
Mitochondrial protein expression in rattus norvegicus and human cells
Confocal microscopy study of the distribution, content and activity of mitochondria during Paracentrotus lividus development
Summary In the present paper we applied confocal microscopy andfluorescence technologies for studying the distribution andthe oxidative activity of sea urchin ( Paracentrotus lividus )mitochondria during development, by in vivo incubating eggsand embryos with cell-permeant MitoTracker probes. Wecalculated, by a mathematical model, the intensity values, the variations of intensity, and the variation index of incorporatedfluorochromes. Data demonstrate that mitochondrial massdoes not change during development, whereas mitochondrialrespirationincreases.Inaddition,startingfrom16blastomeresstage, some regions of the embryo contain organelles moreactive in oxygen consumption. Introduction The con…
EFFECT OF CADMIUM ON IMMORTALIZED AND NEOPLASTIC EPITHELIAL CELLS OF THE HUMAN BREAST
ISOLATION OF CYTOCHROME OXIDASE COMPLEX FROM SEA URCHIN MITOCHONDRIAL INNER MEMBRANES
Cytochrome oxidase complex has been isolated and purified from sea urchin mitochondrial inner membranes. The complex exhibited a discrete number of bands in PAGE and a 17 kDa band was recognized by an anti-human subunit IV antibody. No bands were recognized by an anti-yeast hsp60 antibody and an anti-sea urchin β ATPase subunit.