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Chrystelle Aillaud Christophe Bosc Yasmina Saoudi Eric Denarier Leticia Peris Laila Sago Nicolas Taulet Adeline Cieren Olivia Tort Maria M. Magiera Carsten Janke Virginie Redeker Annie Andrieux Marie-Jo Moutin 《Molecular biology of the cell》2016,27(4):640-653
Cellular α-tubulin can bear various carboxy-terminal sequences: full-length tubulin arising from gene neosynthesis is tyrosinated, and two truncated variants, corresponding to detyrosinated and Δ2 α‑tubulin, result from the sequential cleavage of one or two C-terminal residues, respectively. Here, by using a novel antibody named 3EG that is highly specific to the –EEEG C-terminal sequence, we demonstrate the occurrence in neuronal tissues of a new αΔ3‑tubulin variant corresponding to α1A/B‑tubulin deleted of its last three residues (EEY). αΔ3‑tubulin has a specific distribution pattern: its quantity in the brain is similar to that of αΔ2-tubulin around birth but is much lower in adult tissue. This truncated α1A/B-tubulin variant can be generated from αΔ2-tubulin by the deglutamylases CCP1, CCP4, CCP5, and CCP6 but not by CCP2 and CCP3. Moreover, using 3EG antibody, we identify a C‑terminally truncated β-tubulin form with the same –EEEG C-terminal sequence. Using mass spectrometry, we demonstrate that β2A/B-tubulin is modified by truncation of the four C-terminal residues (EDEA). We show that this newly identified βΔ4-tubulin is ubiquitously present in cells and tissues and that its level is constant throughout the cell cycle. These new C-terminally truncated α- and β-tubulin variants, both ending with –EEEG sequence, are expected to regulate microtubule physiology. Of interest, the αΔ3-tubulin seems to be related to dynamic microtubules, resembling tyrosinated-tubulin rather than the other truncated variants, and may have critical function(s) in neuronal development. 相似文献
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Ablation of PRC1 by small interfering RNA demonstrates that cytokinetic abscission requires a central spindle bundle in mammalian cells, whereas completion of furrowing does not
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Mollinari C Kleman JP Saoudi Y Jablonski SA Perard J Yen TJ Margolis RL 《Molecular biology of the cell》2005,16(3):1043-1055
The temporal and spatial regulation of cytokinesis requires an interaction between the anaphase mitotic spindle and the cell cortex. However, the relative roles of the spindle asters or the central spindle bundle are not clear in mammalian cells. The central spindle normally serves as a platform to localize key regulators of cell cleavage, including passenger proteins. Using time-lapse and immunofluorescence analysis, we have addressed the consequences of eliminating the central spindle by ablation of PRC1, a microtubule bundling protein that is critical to the formation of the central spindle. Without a central spindle, the asters guide the equatorial cortical accumulation of anillin and actin, and of the passenger proteins, which organize into a subcortical ring in anaphase. Furrowing goes to completion, but abscission to create two daughter cells fails. We conclude the central spindle bundle is required for abscission but not for furrowing in mammalian cells. 相似文献
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Kozielski F Skoufias DA Indorato RL Saoudi Y Jungblut PR Hustoft HK Strozynski M Thiede B 《Proteomics》2008,8(2):289-300
Mitotic kinesins represent potential drug targets for anticancer chemotherapy. Inhibitors of different chemical classes have been identified that target human Eg5, a kinesin responsible for the establishment of the bipolar spindle. One potent Eg5 inhibitor is S-trityl-L-cysteine (STLC), which arrests cells in mitosis and exhibits tumor growth inhibition activity. However, the underlying mechanism of STLC action on the molecular level is unknown. Here, cells treated with STLC were blocked in mitosis through activation of the spindle assembly checkpoint as shown by the phosphorylated state of BubR1 and the accumulation of mitosis specific phosphorylation on histone H3 and aurora A kinase. Using live cell imaging, we observed prolonged mitotic arrest and subsequent cell death after incubation of GFP-alpha-tubulin HeLa cells with STLC. Activated caspase-9 occurred before cleavage of caspase-8 leading to the accumulation of the activated executioner caspase-3 suggesting that STLC induces apoptosis through the intrinsic apoptotic pathway. Proteome analysis following STLC treatment revealed 33 differentially regulated proteins of various cellular processes, 31 of which can be linked to apoptotic cell death. Interestingly, four identified proteins, chromobox protein homolog, RNA-binding Src associated in mitosis 68 kDa protein, stathmin, and translationally controlled tumor protein can be linked to mitotic and apoptotic processes. 相似文献
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Saoudi M Abdelmouleh A Kammoun W Ellouze F Jamoussi K El Feki A 《Comptes rendus biologies》2008,331(8):611-616
This study was undertaken to assess the risk of poisoning due to consumption of the puffer fish Lagocephalus lagocephalus collected along the Tunisian coast. Wistar rats were daily intraperitoneally injected, for 10 days, with acidic extracts of liver or flesh (muscles + skin) of L. lagocephalus. Control rats received injections of NaCl (0.9%). No mortality and no evident signs of neurotoxicity were recorded in treated rats. Conversely, treatment led to: (1) diarrhoea and body and organ (liver, kidney) weights loss; (2) oxidative stress evidenced by an increase in lipid peroxidation (TBARS) and conversely a decrease in antioxidant enzyme activities (SOD, catalase, GSH-Px) in tissues (blood cells, liver, kidneys); (3) a decrease in alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities in blood plasma. 相似文献
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A 3-year study assessed the diagnostic accuracy of touch imprint smears in the diagnosis of lung cancer. Touch imprint smears were prepared from 90 computerized tomographic-guided core needle lung biopsies. Cytological diagnosis of touch imprint smears were correlated with the histological diagnosis of the corresponding core needle biopsy specimen, which was taken as the gold standard. The sensitivity, specificity, positive predictive value and negative predictive value of imprint smear results were 89%, 100%, 100% and 68%, respectively. There were no false positives, and all patients with small cell lung cancer were correctly diagnosed with this technique. Imprint cytology can be used to provide a rapid, preliminary diagnosis of lung cancer. 相似文献
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Evidence is accumulating that chromatin plays a major role in the control of cellular response to stress. This is best illustrated by the recent findings that chromatin-modifying factors of class III histone deacetylases (sirtuins) are capable of protecting cells from oxidative and genotoxic stress. In particular, Sirt1 has been shown to mimic the action of caloric restriction for the prevention of aging-associated diseases. In the present study, we have investigated the potential role of class I and II histone deacetylases (HDACs) in cellular protection against various stresses, including those caused by nutrient deprivation. For this, we utilized a cellular model in which expression of class I and II HDACs was altered as a result of cellular adaptation to trichostatin A (TSA), a selective inhibitor of these deacetylases. Our results indicated that TSA-resistant cells also developed resistance to H2 O2 , DNA-damaging agents, and to nutrient deprivation. Interestingly, the insulin signaling pathway mediated by Akt was inhibited in the TSA-resistant cells, mirroring the effect of glucose deprivation on this pathway. Since expression of HDAC4 was consistently enhanced in the TSA-resistant cell lines, we suggest that this enzyme may contribute to their anti-stress response. In agreement with this, siRNA-mediated knockdown of HDAC4 in stress-resistant cells enhanced their sensitivity to the DNA-damaging drug doxorubicin and also to glucose deprivation. Akt phosphorylation was also up-regulated in response to HDC4 knockdown. Together, these findings suggest that cellular conditioning with TSA may represent a useful approach to mimic the effects of caloric restriction. 相似文献
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