70 research outputs found
Evaluation of ground information with respect to EPB tunnelling for the Thessaloniki metro, Greece
Ο Μητροπολιτικός σιδηρόδρομος της Θεσσαλονίκης αποτελείται από δύο παράλληλες σήραγγες διαμέτρου ~6 m και μήκους ~8 km η κάθε μία και περιλαμβάνει 13 σταθμούς. Η γεωλογία του πολεοδομικού συγκροτήματος της Θεσσαλονίκης χαρακτηρίζεται από την παρουσία νεογενών και τεταρτογενών αποθέσεων. Ο κύριος σχηματισμός της περιοχής του έργου είναι μία σειρά πολύ στιφρών έως σκληρών ερυθρών αργίλων ανωμειοκαινικής-πλειοκαινικής ηλικίας. Σχηματισμοί του Τεταρτογενούς που έχουν αποτεθεί πάνω σε αυτές τις αργίλους συνίστανται από αργιλώδεις-ιλυώδεις άμμους ή/και χάλικες. Το πρόγραμμα γεωερευνητικών εργασιών περιελάμβανε έναν σημαντικό αριθμό δειγματοληπτικών γεωτρήσεων, επί τόπου και εργαστηριακών δοκιμών. Τα στοιχεία του γεωερευνητικού προγράμματος αξιολογήθηκαν ώστε να κατανοηθεί καλύτερα το γεωλογικό προσομοίωμα της περιοχής του έργου και να διακριτοποιηθούν ζώνες με βάση τη συμπεριφορά των γεωυλικών κατά τη διάνοιξη της σήραγγας με μηχάνημα ολομέτωπης κοπής (ΤΒΜ). Όσον αφορά το μηχάνημα διάνοιξης, η επιλογή ενός μηχανήματος εδαφικής εξισορροπητικής πίεσης (ΕΡΒΜ) φαίνεται να είναι εύλογη τόσο από πλευράς ευστάθειας όσο και από πλευράς ρυθμού προχώρησης. Η επιλογή αυτή υπαγορεύεται από τα χαρακτηριστικά του εδάφους για την κάλυψη όλων των αντικειμενικών σκοπών όπως ο έλεγχος των καθιζήσεων και εδαφικών μετακινήσεων, η διατήρηση της στάθμης του υπόγειου νερού αλλά και η ικανοποιητική προχώρηση των σηράγγωνThe Thessaloniki Metropolitan Railway comprises two separate ~6 m diameter parallel tunnels with an ~8 km stretch each and 13 stations. The geology of the urban area of Thessaloniki is characterised by the presence of Neogene and Quaternary deposits. The base formation for the project area is a very stiff to hard red clay, dating to Upper Miocene-Pliocene. Upon this formation, Quaternary sediments have been deposited, most of which comprise sand and/or gravel in a clay-silt dominated matrix, covered in places by anthropogenic fill. Ground investigation campaigns incorporated a significant number of sampling boreholes and in situ and laboratory testing. This information was elaborated in order to obtain a better geological understanding and a geotechnical zonation of the ground with respect to mechanized tunnelling. EPB M appears to be the reasonable choice for the project in all aspects of tunnel safety and tunnelling performance. The characteristics and parameters of the soils and the hydrogeological regime directed towards this selection and it is expected that all the objectives, such as settlement and ground movements control, water table level maintenance and adequate performance, will be met by an EPBM provided it is properly operate
Slx8 removes Pli1-dependent protein-SUMO conjugates including SUMOylated Topoisomerase I to promote genome stability
Peer reviewedPublisher PD
SUMO modification of PCNA is controlled by DNA
Post-translational modification by the ubiquitin-like protein SUMO is often regulated by cellular signals that restrict the modification to appropriate situations. Nevertheless, many SUMO-specific ligases do not exhibit much target specificity, and—compared with the diversity of sumoylation substrates—their number is limited. This raises the question of how SUMO conjugation is controlled in vivo. We report here an unexpected mechanism by which sumoylation of the replication clamp protein, PCNA, from budding yeast is effectively coupled to S phase. We find that loading of PCNA onto DNA is a prerequisite for sumoylation in vivo and greatly stimulates modification in vitro. To our surprise, however, DNA binding by the ligase Siz1, responsible for PCNA sumoylation, is not strictly required. Instead, the stimulatory effect of DNA on conjugation is mainly attributable to DNA binding of PCNA itself. These findings imply a change in the properties of PCNA upon loading that enhances its capacity to be sumoylated
The SUMO Isopeptidase Ulp2p Is Required to Prevent Recombination-Induced Chromosome Segregation Lethality following DNA Replication Stress
SUMO conjugation is a key regulator of the cellular response to DNA replication stress, acting in part to control recombination at stalled DNA replication forks. Here we examine recombination-related phenotypes in yeast mutants defective for the SUMO de-conjugating/chain-editing enzyme Ulp2p. We find that spontaneous recombination is elevated in ulp2 strains and that recombination DNA repair is essential for ulp2 survival. In contrast to other SUMO pathway mutants, however, the frequency of spontaneous chromosome rearrangements is markedly reduced in ulp2 strains, and some types of rearrangements arising through recombination can apparently not be tolerated. In investigating the basis for this, we find DNA repair foci do not disassemble in ulp2 cells during recovery from the replication fork-blocking drug methyl methanesulfonate (MMS), corresponding with an accumulation of X-shaped recombination intermediates. ulp2 cells satisfy the DNA damage checkpoint during MMS recovery and commit to chromosome segregation with similar kinetics to wild-type cells. However, sister chromatids fail to disjoin, resulting in abortive chromosome segregation and cell lethality. This chromosome segregation defect can be rescued by overproducing the anti-recombinase Srs2p, indicating that recombination plays an underlying causal role in blocking chromatid separation. Overall, our results are consistent with a role for Ulp2p in preventing the formation of DNA lesions that must be repaired through recombination. At the same time, Ulp2p is also required to either suppress or resolve recombination-induced attachments between sister chromatids. These opposing defects may synergize to greatly increase the toxicity of DNA replication stress
Cdk1 Targets Srs2 to Complete Synthesis-Dependent Strand Annealing and to Promote Recombinational Repair
Cdk1 kinase phosphorylates budding yeast Srs2, a member of UvrD protein family, displays both DNA translocation and DNA unwinding activities in vitro. Srs2 prevents homologous recombination by dismantling Rad51 filaments and is also required for double-strand break (DSB) repair. Here we examine the biological significance of Cdk1-dependent phosphorylation of Srs2, using mutants that constitutively express the phosphorylated or unphosphorylated protein isoforms. We found that Cdk1 targets Srs2 to repair DSB and, in particular, to complete synthesis-dependent strand annealing, likely controlling the disassembly of a D-loop intermediate. Cdk1-dependent phosphorylation controls turnover of Srs2 at the invading strand; and, in absence of this modification, the turnover of Rad51 is not affected. Further analysis of the recombination phenotypes of the srs2 phospho-mutants showed that Srs2 phosphorylation is not required for the removal of toxic Rad51 nucleofilaments, although it is essential for cell survival, when DNA breaks are channeled into homologous recombinational repair. Cdk1-targeted Srs2 displays a PCNA–independent role and appears to have an attenuated ability to inhibit recombination. Finally, the recombination defects of unphosphorylatable Srs2 are primarily due to unscheduled accumulation of the Srs2 protein in a sumoylated form. Thus, the Srs2 anti-recombination function in removing toxic Rad51 filaments is genetically separable from its role in promoting recombinational repair, which depends exclusively on Cdk1-dependent phosphorylation. We suggest that Cdk1 kinase counteracts unscheduled sumoylation of Srs2 and targets Srs2 to dismantle specific DNA structures, such as the D-loops, in a helicase-dependent manner during homologous recombinational repair
The S phase checkpoint promotes the Smc5/6 complex dependent SUMOylation of Pol2, the catalytic subunit of DNA polymerase ε
Replication fork stalling and accumulation of single-stranded DNA trigger the S phase checkpoint, a signalling cascade that, in budding yeast, leads to the activation of the Rad53 kinase. Rad53 is essential in maintaining cell viability, but its targets of regulation are still partially unknown. Here we show that Rad53 drives the hyper-SUMOylation of Pol2, the catalytic subunit of DNA polymerase ε, principally following replication forks stalling induced by nucleotide depletion. Pol2 is the main target of SUMOylation within the replisome and its modification requires the SUMO-ligase Mms21, a subunit of the Smc5/6 complex. Moreover, the Smc5/6 complex co-purifies with Pol ε, independently of other replisome components. Finally, we map Pol2 SUMOylation to a single site within the N-terminal catalytic domain and identify a SUMO-interacting motif at the C-terminus of Pol2. These data suggest that the S phase checkpoint regulate Pol ε during replication stress through Pol2 SUMOylation and SUMO-binding abilit
Genetic predisposition to ductal carcinoma in situ of the breast
Background: Ductal carcinoma in situ (DCIS) is a non-invasive form of breast cancer. It is often associated with invasive ductal carcinoma (IDC), and is considered to be a non-obligate precursor of IDC. It is not clear to what extent these two forms of cancer share low-risk susceptibility loci, or whether there are differences in the strength of association for shared loci. Methods: To identify genetic polymorphisms that predispose to DCIS, we pooled data from 38 studies comprising 5,067 cases of DCIS, 24,584 cases of IDC and 37,467 controls, all genotyped using the iCOGS chip. Results: Most (67 %) of the 76 known breast cancer predisposition loci showed an association with DCIS in the same direction as previously reported for invasive breast cancer. Case-only analysis showed no evidence for differences between associations for IDC and DCIS after considering multiple testing. Analysis by estrogen receptor (ER) status confirmed that loci associated with ER positive IDC were also associated with ER positive DCIS. Analysis of DCIS by grade suggested that two independent SNPs at 11q13.3 near CCND1 were specific to low/intermediate grade DCIS (rs75915166, rs554219). These associations with grade remained after adjusting for ER status and were also found in IDC. We found no novel DCIS-specific loci at a genome wide significance level of P < 5.0x10-8. Conclusion: In conclusion, this study provides the strongest evidence to date of a shared genetic susceptibility for IDC and DCIS. Studies with larger numbers of DCIS are needed to determine if IDC or DCIS specific loci exist
Aristotle’s virtue ethics as a conceptual framework for the study and practice of social work in modern times
Crosstalk between SUMO and Ubiquitin on PCNA Is Mediated by Recruitment of the Helicase Srs2p.
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