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1.
Alexander Mironov Antonino Colanzi Maria Giuseppina Silletta Giusy Fiucci Silvio Flati Aurora Fusella Roman Polishchuk Alexander Mironov Jr. Giuseppe Di Tullio Roberto Weigert Vivek Malhotra Daniela Corda Maria Antonietta De Matteis Alberto Luini 《The Journal of cell biology》1997,139(5):1109-1118
We have investigated the role of the ADP- ribosylation induced by brefeldin A (BFA) in the mechanisms controlling the architecture of the Golgi complex. BFA causes the rapid disassembly of this organelle into a network of tubules, prevents the association of coatomer and other proteins to Golgi membranes, and stimulates the ADP-ribosylation of two cytosolic proteins of 38 and 50 kD (GAPDH and BARS-50; De Matteis, M.A., M. DiGirolamo, A. Colanzi, M. Pallas, G. Di Tullio, L.J. McDonald, J. Moss, G. Santini, S. Bannykh, D. Corda, and A. Luini. 1994. Proc. Natl. Acad. Sci. USA. 91:1114–1118; Di Girolamo, M., M.G. Silletta, M.A. De Matteis, A. Braca, A. Colanzi, D. Pawlak, M.M. Rasenick, A. Luini, and D. Corda. 1995. Proc. Natl. Acad. Sci. USA. 92:7065–7069). To study the role of ADP-ribosylation, this reaction was inhibited by depletion of NAD+ (the ADP-ribose donor) or by using selective pharmacological blockers in permeabilized cells. In NAD+-depleted cells and in the presence of dialized cytosol, BFA detached coat proteins from Golgi membranes with normal potency but failed to alter the organelle's structure. Readdition of NAD+ triggered Golgi disassembly by BFA. This effect of NAD+ was mimicked by the use of pre–ADP- ribosylated cytosol. The further addition of extracts enriched in native BARS-50 abolished the ability of ADP-ribosylated cytosol to support the effect of BFA. Pharmacological blockers of the BFA-dependent ADP-ribosylation (Weigert, R., A. Colanzi, A. Mironov, R. Buccione, C. Cericola, M.G. Sciulli, G. Santini, S. Flati, A. Fusella, J. Donaldson, M. DiGirolamo, D. Corda, M.A. De Matteis, and A. Luini. 1997. J. Biol. Chem. 272:14200–14207) prevented Golgi disassembly by BFA in permeabilized cells. These inhibitors became inactive in the presence of pre–ADP-ribosylated cytosol, and their activity was rescued by supplementing the cytosol with a native BARS-50–enriched fraction. These results indicate that ADP-ribosylation plays a role in the Golgi disassembling activity of BFA, and suggest that the ADP-ribosylated substrates are components of the machinery controlling the structure of the Golgi apparatus. 相似文献
2.
Guiseppina Silletta Maria Colanzi Antonino Weigert Roberto Di Girolamo Maria Santone Ivana Fiucci Giusy Mironov Alexander De Matteis Maria Antonietta Luini Alberto Corda Daniela 《Molecular and cellular biochemistry》1999,193(1-2):43-51
The fungal toxin brefeldin A (BFA) dissociates coat proteins from Golgi membranes, causes the rapid disassembly of the Golgi complex and potently stimulates the ADP-ribosylation of two cytosolic proteins of 38 and 50 kDa. These proteins have been identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and a novel guanine nucleotide binding protein (BARS-50), respectively. The role of ADP-ribosylation in mediating the effects of BFA on the structure and function of the Golgi complex was analyzed by several approaches including the use of selective pharmacological blockers of the reaction and the use of ADP-ribosylated cytosol and/or enriched preparations of the BFA-induced ADP-ribosylation substrates, GAPDH and BARS-50.A series of blockers of the BFA-dependent ADP-ribosylation reaction identified in our laboratory inhibited the effects of BFA on Golgi morphology and, with similar potency, the ADP-ribosylation of BARS-50 and GAPDH. In permeabilized RBL cells, the BFA-dependent disassembly of the Golgi complex required NAD+ and cytosol. Cytosol that had been previously ADP-ribosylated (namely, it contained ADP-ribosylated GAPDH and BARS-50), was instead sufficient to sustain the Golgi disassembly induced by BFA.Taken together, these results indicate that an ADP-ribosylation reaction is part of the mechanism of action of BFA and it might intervene in the control of the structure and function of the Golgi complex. 相似文献
3.
Image processing,diagnostic information extraction and quantitative assessment in pathology 总被引:1,自引:1,他引:0
Santinelli A Mazzucchelli R Colanzi P Tinca A Montironi R 《Journal of cellular and molecular medicine》2002,6(1):93-106
As we enter the information age we hold strong beliefs in the benefits of digital technology applied to pathology: numerical representation offers objectivity . Digital knowledge may indeed lead to significant information discovery, and, processing systems might be designed to allow a true evolution of capabilities. Questions arise whether the methodology underlying quantitative analysis provides the information that we need and whether it is appropriate for some of the problems encountered in diagnostic and prognostic histopathology. While one certainly would not dispute the value of statistical procedures, the clinical needs call for individual patient targeted prognosis. 相似文献
4.
Santinelli A Baccarini M Colanzi P Fabris G 《Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology》2000,22(4):277-284
OBJECTIVE: To study the angiogenic process in intraductal carcinoma of the breast, with and without a small focus of stromal infiltration, and to compare the microvessel density between the in situ phase and the early infiltration phases of breast cancer. STUDY DESIGN: Microvessel density (number of microvessels per square millimeter of neoplasia) was quantitatively evaluated on anti-factor VIII-immunostained histologic sections obtained from 10 ductal carcinomas in situ (DCIS) (category A), 22 DCIS with a small focus of stromal infiltration (category B), 10 microinvasive carcinomas (category C), 12 T1a carcinomas (category D) and 20 T1b carcinomas (category E). RESULTS: The five categories of lesion had different values for microvessel density (P = .0017). Category A had microvessel density lower than category B (P = .0005). Category B had microvessel density higher than categories C, D and E (P = .0028, .0133 and .0033, respectively). CONCLUSION: Microvessel density seems to be a feature related to each crucial step in the early phases of neoplastic progression. 相似文献
5.
Angela Persico Romina Ines Cervigni Maria Luisa Barretta Daniela Corda Antonino Colanzi 《Molecular biology of the cell》2010,21(21):3708-3721
At the onset of mitosis, the Golgi complex undergoes a multistep fragmentation process that is required for its correct partitioning into the daughter cells. Inhibition of this Golgi fragmentation results in cell cycle arrest at the G2 stage, suggesting that correct inheritance of the Golgi complex is monitored by a “Golgi mitotic checkpoint.” However, the molecular basis of this G2 block is not known. Here, we show that the G2-specific Golgi fragmentation stage is concomitant with centrosome recruitment and activation of the mitotic kinase Aurora-A, an essential regulator for entry into mitosis. We show that a block of Golgi partitioning impairs centrosome recruitment and activation of Aurora-A, which results in the G2 block of cell cycle progression. Overexpression of Aurora-A overrides this cell cycle block, indicating that Aurora-A is a major effector of the Golgi checkpoint. Our findings provide the basis for further understanding of the signaling pathways that coordinate organelle inheritance and cell duplication. 相似文献
6.
Bonazzi M Spanò S Turacchio G Cericola C Valente C Colanzi A Kweon HS Hsu VW Polishchuck EV Polishchuck RS Sallese M Pulvirenti T Corda D Luini A 《Nature cell biology》2005,7(6):570-580
Membrane fission is a fundamental step in membrane transport. So far, the only fission protein machinery that has been implicated in in vivo transport involves dynamin, and functions in several, but not all, transport pathways. Thus, other fission machineries may exist. Here, we report that carboxy-terminal binding protein 3/brefeldin A-ribosylated substrate (CtBP3/BARS) controls fission in basolateral transport from the Golgi to the plasma membrane and in fluid-phase endocytosis, whereas dynamin is not involved in these steps. Conversely, CtBP3/BARS protein is inactive in apical transport to the plasma membrane and in receptor-mediated endocytosis, both steps being controlled by dynamin. This indicates that CtBP3/BARS controls membrane fission in endocytic and exocytic transport pathways, distinct from those that require dynamin. 相似文献
7.
Golgi complex fragmentation in G2/M transition: An organelle-based cell-cycle checkpoint 总被引:1,自引:0,他引:1
In mammalian cells, the Golgi complex is organized into a continuous membranous system known as the Golgi ribbon, which is formed by individual Golgi stacks that are laterally connected by tubular bridges. During mitosis, the Golgi ribbon undergoes extensive fragmentation through a multistage process that is required for its correct partitioning into the daughter cells. Importantly, inhibition of this Golgi disassembly results in cell-cycle arrest at the G2 stage, suggesting that accurate inheritance of the Golgi complex is monitored by a "Golgi mitotic checkpoint." Here, we discuss the mechanisms and regulation of the Golgi ribbon breakdown and briefly comment on how Golgi partitioning may inhibit G2/M transition. 相似文献
8.
Wilson C Venditti R Rega LR Colanzi A D'Angelo G De Matteis MA 《The Biochemical journal》2011,433(1):1-9
Remarkable advances have been made during the last few decades in defining the organizational principles of the secretory pathway. The Golgi complex in particular has attracted special attention due to its central position in the pathway, as well as for its fascinating and complex structure. Analytical studies of this organelle have produced significant advances in our understanding of its function, although some aspects still seem to elude our comprehension. In more recent years a level of complexity surrounding this organelle has emerged with the discovery that the Golgi complex is involved in cellular processes other than the 'classical' trafficking and biosynthetic pathways. The resulting picture is that the Golgi complex can be considered as a cellular headquarters where cargo sorting/processing, basic metabolism, signalling and cell-fate decisional processes converge. 相似文献
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10.
The Golgi membranes, in the form of stacks of cisternae, are contained in the pericentriolar region of mammalian cells. During mitosis, these membranes are fragmented and dispersed throughout the cell. Recent studies are revealing the significance of pericentriolar position of the Golgi apparatus and mechanism by which these membranes are fragmented during mitosis. 相似文献