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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd.
    Molecular microbiology 43 (2002), S. 0 
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: In Escherichia coli, UV-irradiated cells resume DNA synthesis after a transient inhibition by a process called replication restart. To elucidate the role of several key proteins involved in this process, we have analysed the time dependence of replication restart in strains carrying a combination of mutations in lexA, recA, polB (pol II), umuDC (pol V), priA, dnaC, recF, recO or recR. We find that both pol II and the origin-independent primosome-assembling function of PriA are essential for the immediate recovery of DNA synthesis after UV irradiation. In their absence, translesion replication or ‘replication readthrough’ occurs ≈ 50 min after UV and is pol V-dependent. In a wild-type, lexA+ background, mutations in recF, recO or recR block both pathways. Similar results were obtained with a lexA(Def) recF strain. However, lexA(Def) recO or lexA(Def) recR strains, although unable to facilitate PriA-pol II-dependent restart, were able to perform pol V-dependent readthrough. The defects in restart attributed to mutations in recF, recO or recR were suppressed in a recA730 lexA(Def) strain expressing constitutively activated RecA (RecA*). Our data suggest that in a wild-type background, RecF, O and R are important for the induction of the SOS response and the formation of RecA*-dependent recombination intermediates necessary for PriA/Pol II-dependent replication restart. In con-trast, only RecF is required for the activation of RecA that leads to the formation of pol V (UmuD′2C) and facilitates replication readthrough.
    Type of Medium: Electronic Resource
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  • 2
    Publication Date: 2018-11-29
    Description: Key Points MYC mediated upregulation of TPX2, KPNA2 and RanGAP1 dysregulate the spindle assembly checkpoint in drug-induced polyploid cells. Drug-induced polyploid cells re-enter cell cycle via multipolar mitosis, fission or budding, a mechanism of disease relapse. Abstract Double-hit (DH) or double-expresser (DE) diffuse large B-cell lymphomas (DLBCL) are high-grade B-cell lymphomas that are mostly incurable with standard chemo-immunotherapy due to treatment resistance. The generation of drug-induced aneuploid/polyploid (DIAP) cells is a common effect of anti-DLBCL therapies (e.g. vincristine, doxorubicin). DIAP cells are thought to be responsible for treatment resistance, as they are capable of re-entering the cell cycle during off-therapy periods. Previously (DOI: 10.1158/1535-7163.MCT-17-0089) we demonstrated that combination of alisertib plus ibrutinib plus rituximab can partially abrogate DIAP cells and induce cell death. Here, we provide evidence that DIAP cells can re-enter the cell cycle and escape apoptosis during anti-DLBCL treatment. We also discuss MYC/BCL2 mediated molecular mechanisms that underlie treatment resistance. We isolated aneuploid/polyploid populations (2n-similar, 4n and 8n) of DH/DE-DLBCL cells after treatment with the aurora kinase (AK) inhibitor alisertib. Time-lapse microscopy of single polyploid cells (8n) revealed that following drug removal, these cells divide and proliferate by reductive cell divisions, including multipolar mitosis, meiosis-like nuclear fission and budding. Genomic, proteomic, and kinomic profiling of DIAP cells demonstrated up-regulation DNA damage, DNA replication and immune evasion pathways. In addition, the immune-kinome, AKT/MTorc1/2, and ERK/MAPK pathway are over-represented in 8n cells. Further, MYC-mediates dysregulation of the spindle assembly checkpoints by over-expression of RanGAP1, TPX2 and KPNA2. Multiple mechanisms contribute to DIAP which are amenable to novel therapeutic intervention(s) in MYC/BCL2-addicted high-grade B-cell lymphomas. Disclosures No relevant conflicts of interest to declare.
    Print ISSN: 0006-4971
    Electronic ISSN: 1528-0020
    Topics: Biology , Medicine
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