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    Highly synchronous mitotic progression in schizosaccharomyces pombe upon relief of transient Cdc2-asM17 inhibition

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    Authors
    Singh, Pawan
    Halova, Lenka
    Hagan, Iain M
    Affiliation
    Cell Division Group, CRUK Manchester Institute, The University of Manchester, Alderley Park, UK.
    Issue Date
    2021
    
    Metadata
    Show full item record
    Abstract
    Synchronized progression of a cell population through the cell division cycle supports the biochemical and functional dissection of cell cycle controls and execution. The concerted behaviour of the population reflects the attributes of each cell within that population. The reversible imposition of a block to cell cycle progression at the G2-M boundary through transient inactivation of the Cdk1-Cyclin B activating phosphatase, Cdc25, with the temperature sensitive cdc25-22 mutant, has been widely used to study fission yeast mitosis and DNA replication. However, the biology of the compromised Cdc25-22 phosphatase generates significant division abnormalities upon release from mitotic arrest. We show how reversible inhibition of Cdc2-asM17, with the ATP analog 3-BrB-PP1, generates higher levels of synchrony with timing and morphology much more reminiscent of a normal division. We also describe a version of the H1 kinase assay of Cdk1-Cyclin B activity that is widely used to monitor mitotic progression which does not require radiolabeled ATP.
    Citation
    Singh P, Halova L, Hagan IM. Highly Synchronous Mitotic Progression in Schizosaccharomyces pombe Upon Relief of Transient Inhibition. In: Methods in Molecular Biology . Springer US; 2021. p. 123–42. 
    Journal
    Methods in Molecular Biology
    URI
    http://hdl.handle.net/10541/624257
    DOI
    10.1007/978-1-0716-1538-6_10
    PubMed ID
    34085220
    Additional Links
    https://dx.doi.org/10.1007/978-1-0716-1538-6_10
    Type
    Article
    Language
    en
    ae974a485f413a2113503eed53cd6c53
    10.1007/978-1-0716-1538-6_10
    Scopus Count
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    All Paterson Institute for Cancer Research

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