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  • 1
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    Temporal regulation of the Mus81-Mms4 endonuclease ensures cell survival under conditions of DNA damage (2013)
    Oxford University Press
    Details
    Publication Date: 2013-10-19
    Description: The structure-specific Mus81-Eme1/Mms4 endonuclease contributes importantly to DNA repair and genome integrity maintenance. Here, using budding yeast, we have studied its function and regulation during the cellular response to DNA damage and show that this endonuclease is necessary for successful chromosome replication and cell survival in the presence of DNA lesions that interfere with replication fork progression. On the contrary, Mus81-Mms4 is not required for coping with replicative stress originated by acute treatment with hydroxyurea (HU), which causes fork stalling. Despite its requirement for dealing with DNA lesions that hinder DNA replication, Mus81-Mms4 activation is not induced by DNA damage at replication forks. Full Mus81-Mms4 activity is only acquired when cells finish S-phase and the endonuclease executes its function after the bulk of genome replication is completed. This post-replicative mode of action of Mus81-Mms4 limits its nucleolytic activity during S-phase, thus avoiding the potential cleavage of DNA substrates that could cause genomic instability during DNA replication. At the same time, it constitutes an efficient fail-safe mechanism for processing DNA intermediates that cannot be resolved by other proteins and persist after bulk DNA synthesis, which guarantees the completion of DNA repair and faithful chromosome replication when the DNA is damaged.
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
    Published by Oxford University Press
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  • 2
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    Uninterrupted MCM2-7 function required for DNA replication fork progression (2000)
    American Association for the Advancement of Science (AAAS)
    Details
    Publication Date: 2000-06-02
    Description: Little is known about the DNA helicases required for the elongation phase of eukaryotic chromosome replication. Minichromosome maintenance (MCM) protein complexes have DNA helicase activity but have only been functionally implicated in initiating DNA replication. Using an improved method for constructing conditional degron mutants, we show that depletion of MCMs after initiation irreversibly blocks the progression of replication forks in Saccharomyces cerevisiae. Like the Escherichia coli dnaB and SV40 T antigen helicases, therefore, the MCM complex is loaded at origins before initiation and is essential for elongation. Restricting MCM loading to the G(1) phase ensures that initiation and elongation occur just once per cell cycle.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Labib, K -- Tercero, J A -- Diffley, J F -- New York, N.Y. -- Science. 2000 Jun 2;288(5471):1643-7.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Imperial Cancer Research Fund Clare Hall Laboratories, South Mimms, Hertfordshire, EN6 3LD, UK.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/10834843" target="_blank"〉PubMed〈/a〉
    Keywords: Cell Cycle Proteins/genetics/*metabolism ; Chromosomal Proteins, Non-Histone ; Chromosomes, Fungal/metabolism ; *DNA Replication ; DNA, Fungal/biosynthesis ; *DNA-Binding Proteins ; Fungal Proteins/genetics/*metabolism ; G1 Phase ; G2 Phase ; *Ligases ; Minichromosome Maintenance Complex Component 4 ; Minichromosome Maintenance Complex Component 6 ; Mitosis ; Mutation ; Recombinant Fusion Proteins/metabolism ; Replicon ; S Phase ; Saccharomyces cerevisiae/genetics/*metabolism ; *Saccharomyces cerevisiae Proteins ; Temperature ; *Ubiquitin-Protein Ligases
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
    Published by American Association for the Advancement of Science (AAAS)
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  • 3
    Electronic Resource
    Electronic Resource
    Encoding genes for endosperm proteins in Hordeum chilense (1991)
    Springer
    Theoretical and applied genetics 81 (1991), S. 127-132 
    Details
    ISSN: 1432-2242
    Keywords: Hordeum ; A-PAGE ; SDS-PAGE ; Two-dimensional electrophoresis ; Hordeins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The endosperm proteins encoded by the genome Hch in Hordeum chilense, Tritordeum (amphiploid Hordeum chilense x Triticum turgidum), common wheat-H. chilense addition lines, and the segregating plants resulting from the cross Tritordeum x T. turgidum, were fractionated by three electrophoretical techniques: SDS-PAGE, A-PAGE, and bidimensional PAGE. Prolamin subunits with a high molecular weight (HMW) were well visualized by SDS-PAGE, the A-PAGE technique permitted good resolution for many hordeins and gliadins, and two-dimensional electrophoresis allowed new sets of bands coded by gene complexes from H. chilense chromosomes to be distinguished. The loci Hor-Hch1 (up to 11 subunits belonging to the ω-, β — and α-hordeins), Glu-Hch1 (one HMW prolamin subunit), Hor-Hch2 (one α-hordein), and Hor-Hch3 (up to four α-hordeins) were located on the H. chilense chromosomes 1Hch, 5Hch, and 7Hch.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00226122
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  • 4
    Electronic Resource
    Electronic Resource
    StgR, a new Streptomyces alboniger member of the LysR family of transcriptional regulators (1998)
    Springer
    Molecular genetics and genomics 259 (1998), S. 475-483 
    Details
    ISSN: 1617-4623
    Keywords: Key wordsStreptomyces ; LysR-type repressor ; stg gene repression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 3240-bp DNA fragment, located next to the puromycin biosynthetic gene cluster of Streptomycesalboniger, contains three complete ORFs in the order: stgA, stgU and stgR. The transcriptional orientation of stgA is opposite to that of stgU and stgR. Each gene is expressed from its own promoter, although stgU and stgR can be cotranscribed. The deduced amino acid sequences of their products present similarities to a variety of pyridoxal-phosphate-dependent aspartate aminotransferases (StgA), several proteins of unknown function (StgU), and the LysR-type of transcriptional regulators (StgR). In a ΔstgR null mutant of S. alboniger, transcription of stgA and stgU is increased with respect to that in the wild type. In addition, in vivo experiments with promoter-probe plasmids indicated that in the ΔstgR mutant, stgA- or stgU-promoter-dependent expression of the reporter gene was up to three-fold higher than in the wild type. Taken together, these results indicate that StgR is a LysR-type transcriptional repressor of both stgA and stgU.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380050838
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  • 5
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    Cell cycle-dependent regulation of the nuclease activity of Mus81-Eme1/Mms4 (2012)
    Oxford University Press
    Details
    Publication Date: 2012-09-27
    Description: The conserved heterodimeric endonuclease Mus81–Eme1/Mms4 plays an important role in the maintenance of genomic integrity in eukaryotic cells. Here, we show that budding yeast Mus81–Mms4 is strictly regulated during the mitotic cell cycle by Cdc28 (CDK)- and Cdc5 (Polo-like kinase)-dependent phosphorylation of the non-catalytic subunit Mms4. The phosphorylation of this protein occurs only after bulk DNA synthesis and before chromosome segregation, and is absolutely necessary for the function of the Mus81–Mms4 complex. Consistently, a phosphorylation-defective mms4 mutant shows highly reduced nuclease activity and increases the sensitivity of cells lacking the RecQ-helicase Sgs1 to various agents that cause DNA damage or replicative stress. The mode of regulation of Mus81–Mms4 restricts its activity to a short period of the cell cycle, thus preventing its function during chromosome replication and the negative consequences for genome stability derived from its nucleolytic action. Yet, the controlled Mus81–Mms4 activity provides a safeguard mechanism to resolve DNA intermediates that may remain after replication and require processing before mitosis.
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
    Published by Oxford University Press
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  • 6
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    DNA synthesis at individual replication forks requires the essential initiation factor Cdc45p (2000)
    EMBO Press
    Details
    Publication Date: 2000-05-02
    Print ISSN: 0261-4189
    Electronic ISSN: 1460-2075
    Topics: Biology , Medicine
    Published by EMBO Press on behalf of European Molecular Biology Organization.
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