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  1. Article: Using an ImageJ-based script to detect replication stress and associated cell cycle exit from G2 phase by fluorescence microscopy.

    Lindqvist, Arne / Hao, Zhiyu / Akopyan, Karen

    Methods in cell biology

    2023  Volume 182, Page(s) 187–197

    Abstract: Replication stress risks genomic integrity. Depending on the level, replication stress can lead to slower progression through S phase and entry into G2 phase with DNA damage. In G2 phase, cells either recover and eventually enter mitosis or permanently ... ...

    Abstract Replication stress risks genomic integrity. Depending on the level, replication stress can lead to slower progression through S phase and entry into G2 phase with DNA damage. In G2 phase, cells either recover and eventually enter mitosis or permanently withdraw from the cell cycle. Here we describe a method to detect cell cycle distribution, replication stress and cell cycle exit from G2 phase using fluorescence microscopy. We provide a script to automate the analysis using ImageJ. The focus has been to make a script and setup that is accessible to people without extensive computer knowledge.
    MeSH term(s) Humans ; Cell Cycle/genetics ; G2 Phase ; Mitosis ; DNA Damage ; Microscopy, Fluorescence ; DNA Replication
    Language English
    Publishing date 2023-03-25
    Publishing country United States
    Document type Journal Article
    ISSN 0091-679X
    ISSN 0091-679X
    DOI 10.1016/bs.mcb.2023.02.003
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: DNA replication and mitotic entry: A brake model for cell cycle progression.

    Lemmens, Bennie / Lindqvist, Arne

    The Journal of cell biology

    2019  Volume 218, Issue 12, Page(s) 3892–3902

    Abstract: The core function of the cell cycle is to duplicate the genome and divide the duplicated DNA into two daughter cells. These processes need to be carefully coordinated, as cell division before DNA replication is complete leads to genome instability and ... ...

    Abstract The core function of the cell cycle is to duplicate the genome and divide the duplicated DNA into two daughter cells. These processes need to be carefully coordinated, as cell division before DNA replication is complete leads to genome instability and cell death. Recent observations show that DNA replication, far from being only a consequence of cell cycle progression, plays a key role in coordinating cell cycle activities. DNA replication, through checkpoint kinase signaling, restricts the activity of cyclin-dependent kinases (CDKs) that promote cell division. The S/G2 transition is therefore emerging as a crucial regulatory step to determine the timing of mitosis. Here we discuss recent observations that redefine the coupling between DNA replication and cell division and incorporate these insights into an updated cell cycle model for human cells. We propose a cell cycle model based on a single trigger and sequential releases of three molecular brakes that determine the kinetics of CDK activation.
    MeSH term(s) Animals ; Cell Cycle Checkpoints ; Cell Cycle Proteins/metabolism ; Cell Division ; Cell Proliferation ; Cyclin-Dependent Kinases/metabolism ; DNA Damage ; DNA Replication ; Enzyme Activation ; Humans ; Kinetics ; Mice ; Mitosis ; Signal Transduction
    Chemical Substances Cell Cycle Proteins ; Cyclin-Dependent Kinases (EC 2.7.11.22)
    Language English
    Publishing date 2019-11-11
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 218154-x
    ISSN 1540-8140 ; 0021-9525
    ISSN (online) 1540-8140
    ISSN 0021-9525
    DOI 10.1083/jcb.201909032
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery.

    Lafranchi, Lorenzo / Müllers, Erik / Rutishauser, Dorothea / Lindqvist, Arne

    Cells

    2020  Volume 9, Issue 9

    Abstract: Cells recovering from the G2/M DNA damage checkpoint rely more on Aurora A-PLK1 signaling than cells progressing through an unperturbed G2 phase, but the reason for this discrepancy is not known. Here, we devised a method based on a FRET reporter for ... ...

    Abstract Cells recovering from the G2/M DNA damage checkpoint rely more on Aurora A-PLK1 signaling than cells progressing through an unperturbed G2 phase, but the reason for this discrepancy is not known. Here, we devised a method based on a FRET reporter for PLK1 activity to sort cells in distinct populations within G2 phase. We employed mass spectroscopy to characterize changes in protein levels through an unperturbed G2 phase and validated that ATAD2 levels decrease in a proteasome-dependent manner. Comparing unperturbed cells with cells recovering from DNA damage, we note that at similar PLK1 activities, recovering cells contain higher levels of Cyclin B1 and increased phosphorylation of CDK1 targets. The increased Cyclin B1 levels are due to continuous Cyclin B1 production during a DNA damage response and are sustained until mitosis. Whereas partial inhibition of PLK1 suppresses mitotic entry more efficiently when cells recover from a checkpoint, partial inhibition of CDK1 suppresses mitotic entry more efficiently in unperturbed cells. Our findings provide a resource for proteome changes during G2 phase, show that the mitotic entry network is rewired during a DNA damage response, and suggest that the bottleneck for mitotic entry shifts from CDK1 to PLK1 after DNA damage.
    MeSH term(s) ATPases Associated with Diverse Cellular Activities/genetics ; ATPases Associated with Diverse Cellular Activities/metabolism ; Aurora Kinase A/genetics ; Aurora Kinase A/metabolism ; CDC2 Protein Kinase/genetics ; CDC2 Protein Kinase/metabolism ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/metabolism ; Cell Line, Tumor ; Cyclin B1/genetics ; Cyclin B1/metabolism ; DNA Damage ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Fibroblasts/cytology ; Fibroblasts/drug effects ; Fibroblasts/metabolism ; Flow Cytometry ; Fluorescence Resonance Energy Transfer ; G2 Phase Cell Cycle Checkpoints/drug effects ; G2 Phase Cell Cycle Checkpoints/genetics ; Gene Expression Regulation ; Humans ; M Phase Cell Cycle Checkpoints/drug effects ; M Phase Cell Cycle Checkpoints/genetics ; Mitosis/drug effects ; Phosphorylation ; Protein Serine-Threonine Kinases/genetics ; Protein Serine-Threonine Kinases/metabolism ; Proto-Oncogene Proteins/genetics ; Proto-Oncogene Proteins/metabolism ; Signal Transduction ; Tumor Suppressor p53-Binding Protein 1/genetics ; Tumor Suppressor p53-Binding Protein 1/metabolism ; Zinostatin/pharmacology ; Polo-Like Kinase 1
    Chemical Substances CCNB1 protein, human ; Cell Cycle Proteins ; Cyclin B1 ; DNA-Binding Proteins ; Proto-Oncogene Proteins ; TP53BP1 protein, human ; Tumor Suppressor p53-Binding Protein 1 ; Zinostatin (9014-02-2) ; AURKA protein, human (EC 2.7.11.1) ; Aurora Kinase A (EC 2.7.11.1) ; Protein Serine-Threonine Kinases (EC 2.7.11.1) ; CDC2 Protein Kinase (EC 2.7.11.22) ; CDK1 protein, human (EC 2.7.11.22) ; ATAD2 protein, human (EC 3.6.1.3) ; ATPases Associated with Diverse Cellular Activities (EC 3.6.4.-)
    Language English
    Publishing date 2020-09-19
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells9092126
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Electron Kinetic Entropy across Quasi-Perpendicular Shocks.

    Lindberg, Martin / Vaivads, Andris / Raptis, Savvas / Lindqvist, Per-Arne / Giles, Barbara L / Gershman, Daniel Jonathan

    Entropy (Basel, Switzerland)

    2022  Volume 24, Issue 6

    Abstract: We use Magnetospheric Multiscale (MMS) data to study electron kinetic entropy per particle Se across Earth's quasi-perpendicular bow shock. We have selected 22 shock crossings covering a wide range of shock conditions. Measured distribution functions are ...

    Abstract We use Magnetospheric Multiscale (MMS) data to study electron kinetic entropy per particle Se across Earth's quasi-perpendicular bow shock. We have selected 22 shock crossings covering a wide range of shock conditions. Measured distribution functions are calibrated and corrected for spacecraft potential, secondary electron contamination, lack of measurements at the lowest energies and electron density measurements based on plasma frequency measurements. All crossings display an increase in electron kinetic entropy across the shock ΔSe being positive or zero within their error margin. There is a strong dependence of ΔSe on the change in electron temperature, ΔTe, and the upstream electron plasma beta, βe. Shocks with large ΔTe have large ΔSe. Shocks with smaller βe are associated with larger ΔSe. We use the values of ΔSe, ΔTe and density change Δne to determine the effective adiabatic index of electrons for each shock crossing. The average effective adiabatic index is ⟨γe⟩=1.64±0.07.
    Language English
    Publishing date 2022-05-24
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2014734-X
    ISSN 1099-4300 ; 1099-4300
    ISSN (online) 1099-4300
    ISSN 1099-4300
    DOI 10.3390/e24060745
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

    Lorenzo Lafranchi / Erik Müllers / Dorothea Rutishauser / Arne Lindqvist

    Cells, Vol 9, Iss 2126, p

    2020  Volume 2126

    Abstract: Cells recovering from the G2/M DNA damage checkpoint rely more on Aurora A-PLK1 signaling than cells progressing through an unperturbed G2 phase, but the reason for this discrepancy is not known. Here, we devised a method based on a FRET reporter for ... ...

    Abstract Cells recovering from the G2/M DNA damage checkpoint rely more on Aurora A-PLK1 signaling than cells progressing through an unperturbed G2 phase, but the reason for this discrepancy is not known. Here, we devised a method based on a FRET reporter for PLK1 activity to sort cells in distinct populations within G2 phase. We employed mass spectroscopy to characterize changes in protein levels through an unperturbed G2 phase and validated that ATAD2 levels decrease in a proteasome-dependent manner. Comparing unperturbed cells with cells recovering from DNA damage, we note that at similar PLK1 activities, recovering cells contain higher levels of Cyclin B1 and increased phosphorylation of CDK1 targets. The increased Cyclin B1 levels are due to continuous Cyclin B1 production during a DNA damage response and are sustained until mitosis. Whereas partial inhibition of PLK1 suppresses mitotic entry more efficiently when cells recover from a checkpoint, partial inhibition of CDK1 suppresses mitotic entry more efficiently in unperturbed cells. Our findings provide a resource for proteome changes during G2 phase, show that the mitotic entry network is rewired during a DNA damage response, and suggest that the bottleneck for mitotic entry shifts from CDK1 to PLK1 after DNA damage.
    Keywords G2 ; mitotic entry ; checkpoint recovery ; FRET ; PLK1 ; Cyclin B1 ; Biology (General) ; QH301-705.5
    Subject code 612
    Language English
    Publishing date 2020-09-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: Cyclin B-Cdk1 activates its own pump to get into the nucleus.

    Lindqvist, Arne

    The Journal of cell biology

    2010  Volume 189, Issue 2, Page(s) 197–199

    Abstract: The transition to mitosis requires extensive nuclear and cytoplasmic rearrangements that must be spatially and temporally coordinated. In this issue, Gavet and Pines (2010a. J. Cell Biol. doi:10.1083/jcb.200909144) report on a simple yet elegant ... ...

    Abstract The transition to mitosis requires extensive nuclear and cytoplasmic rearrangements that must be spatially and temporally coordinated. In this issue, Gavet and Pines (2010a. J. Cell Biol. doi:10.1083/jcb.200909144) report on a simple yet elegant mechanism as to how this is achieved. By monitoring the activity of cyclin B-Cdk1 in real time, the authors show that concomitant with its activation in the cytoplasm, the kinase complex is rapidly imported into the nucleus by modifying the activity of the nucleocytoplasmic transport machinery. Thus, cyclin B-Cdk1 activates its own pump to get into the nucleus.
    MeSH term(s) Active Transport, Cell Nucleus/physiology ; CDC2 Protein Kinase/genetics ; CDC2 Protein Kinase/metabolism ; Cell Cycle/physiology ; Cell Nucleus/metabolism ; Chromatin/metabolism ; Cyclin B/genetics ; Cyclin B/metabolism ; Cytoplasm/metabolism ; Enzyme Activation
    Chemical Substances Chromatin ; Cyclin B ; CDC2 Protein Kinase (EC 2.7.11.22)
    Language English
    Publishing date 2010-04-19
    Publishing country United States
    Document type Comment ; Journal Article
    ZDB-ID 218154-x
    ISSN 1540-8140 ; 0021-9525
    ISSN (online) 1540-8140
    ISSN 0021-9525
    DOI 10.1083/jcb.201003032
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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  7. Article ; Online: How the cell cycle enforces senescence.

    Silva Cascales, Helena / Müllers, Erik / Lindqvist, Arne

    Aging

    2017  Volume 9, Issue 10, Page(s) 2022–2023

    MeSH term(s) Animals ; Cell Cycle/physiology ; Cellular Senescence/physiology ; DNA Damage ; Gene Expression Regulation ; Genomic Instability
    Language English
    Publishing date 2017-10-29
    Publishing country United States
    Document type Journal Article
    ISSN 1945-4589
    ISSN (online) 1945-4589
    DOI 10.18632/aging.101316
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  8. Article ; Online: Author Correction: Downstream high-speed plasma jet generation as a direct consequence of shock reformation.

    Raptis, Savvas / Karlsson, Tomas / Vaivads, Andris / Pollock, Craig / Plaschke, Ferdinand / Johlander, Andreas / Trollvik, Henriette / Lindqvist, Per-Arne

    Nature communications

    2022  Volume 13, Issue 1, Page(s) 986

    Language English
    Publishing date 2022-02-16
    Publishing country England
    Document type Published Erratum
    ZDB-ID 2553671-0
    ISSN 2041-1723 ; 2041-1723
    ISSN (online) 2041-1723
    ISSN 2041-1723
    DOI 10.1038/s41467-022-28664-3
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article: Bystander communication and cell cycle decisions after DNA damage.

    Jaiswal, Himjyot / Lindqvist, Arne

    Frontiers in genetics

    2015  Volume 6, Page(s) 63

    Abstract: The DNA damage response (DDR) has two main goals, to repair the damaged DNA and to communicate the presence of damaged DNA. This communication allows the adaptation of cellular behavior to minimize the risk associated with DNA damage. In particular, cell ...

    Abstract The DNA damage response (DDR) has two main goals, to repair the damaged DNA and to communicate the presence of damaged DNA. This communication allows the adaptation of cellular behavior to minimize the risk associated with DNA damage. In particular, cell cycle progression must be adapted after a DNA-damaging insult, and cells either pause or terminally exit the cell cycle during a DDR. As cells can accumulate mutations after a DDR due to error-prone DNA repair, terminal cell cycle exit may prevent malignant transformation. The tumor suppressor p53 plays a key role in promoting terminal cell cycle exit. Interestingly, p53 has been implicated in communication of a stress response to surrounding cells, known as the bystander response. Recently, surrounding cells have also been shown to affect the damaged cell, suggesting the presence of intercellular feedback loops. How such feedback may affect terminal cell cycle exit remains unclear, but its presence calls for caution in evaluating cellular outcome without controlling the cellular surrounding. In addition, such feedback may contribute to how the cellular environment affects malignant transformation after DNA damage.
    Language English
    Publishing date 2015-02-27
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2606823-0
    ISSN 1664-8021
    ISSN 1664-8021
    DOI 10.3389/fgene.2015.00063
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Electron scale coherent structure as micro accelerator in the Earth's magnetosheath.

    Xie, Zi-Kang / Zong, Qiu-Gang / Yue, Chao / Zhou, Xu-Zhi / Liu, Zhi-Yang / He, Jian-Sen / Hao, Yi-Xin / Ng, Chung-Sang / Zhang, Hui / Yao, Shu-Tao / Pollock, Craig / Le, Guan / Ergun, Robert / Lindqvist, Per-Arne

    Nature communications

    2024  Volume 15, Issue 1, Page(s) 886

    Abstract: Turbulent energy dissipation is a fundamental process in plasma physics that has not been settled. It is generally believed that the turbulent energy is dissipated at electron scales leading to electron energization in magnetized plasmas. Here, we ... ...

    Abstract Turbulent energy dissipation is a fundamental process in plasma physics that has not been settled. It is generally believed that the turbulent energy is dissipated at electron scales leading to electron energization in magnetized plasmas. Here, we propose a micro accelerator which could transform electrons from isotropic distribution to trapped, and then to stream (Strahl) distribution. From the MMS observations of an electron-scale coherent structure in the dayside magnetosheath, we identify an electron flux enhancement region in this structure collocated with an increase of magnetic field strength, which is also closely associated with a non-zero parallel electric field. We propose a trapping model considering a field-aligned electric potential together with the mirror force. The results are consistent with the observed electron fluxes from ~50 eV to ~200 eV. It further demonstrates that bidirectional electron jets can be formed by the hourglass-like magnetic configuration of the structure.
    Language English
    Publishing date 2024-01-30
    Publishing country England
    Document type Journal Article
    ZDB-ID 2553671-0
    ISSN 2041-1723 ; 2041-1723
    ISSN (online) 2041-1723
    ISSN 2041-1723
    DOI 10.1038/s41467-024-45040-5
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