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    Interaction of Proliferating Cell Nuclear Antigen With DNA at the Single Molecule Level

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    Name:
    Vlad Stefan Raducanu Thesis.pdf
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    2.037Mb
    Format:
    PDF
    Description:
    Vlad-Stefan Raducanu Final Thesis
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    Type
    Thesis
    Authors
    Raducanu, Vlad-Stefan cc
    Advisors
    Hamdan, Samir cc
    Committee members
    Habuchi, Satoshi cc
    Liberale, Carlo cc
    Program
    Bioscience
    KAUST Department
    Biological and Environmental Sciences and Engineering (BESE) Division
    Date
    2016-05
    Permanent link to this record
    http://hdl.handle.net/10754/609052
    
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    Abstract
    Proliferating cell nuclear antigen (PCNA) is a key factor involved in Eukaryotic DNA replication and repair, as well as other cellular pathways. Its importance comes mainly from two aspects: the large numbers of interacting partners and the mechanism of facilitated diffusion along the DNA. The large numbers of interacting partners makes PCNA a necessary factor to consider when studying DNA replication, either in vitro or in vivo. The mechanism of facilitated diffusion along the DNA, i.e. sliding along the duplex, reduces the six degrees of freedom of the molecule, three degrees of freedom of translation and three degrees of freedom of rotation, to only two, translation along the duplex and rotational tracking of the helix. Through this mechanism PCNA can recruit its partner proteins and localize them to the right spot on the DNA, maybe in the right spatial orientation, more effectively and in coordination with other proteins. Passive loading of the closed PCNA ring on the DNA without free ends is a topologically forbidden process. Replication factor C (RFC) uses energy of ATP hydrolysis to mechanically open the PCNA ring and load it on the dsDNA. The first half of the introduction gives overview of PCNA and RFC and the loading mechanism of PCNA on dsDNA. The second half is dedicated to a diffusion model and to an algorithm for analyzing PCNA sliding. PCNA and RFC were successfully purified, simulations and a mean squared displacement analysis algorithm were run and showed good stability and experimental PCNA sliding data was analyzed and led to parameters similar to the ones in literature.
    Citation
    Raducanu, V.-S. (2016). Interaction of Proliferating Cell Nuclear Antigen With DNA at the Single Molecule Level. KAUST Research Repository. https://doi.org/10.25781/KAUST-I4JFX
    DOI
    10.25781/KAUST-I4JFX
    ae974a485f413a2113503eed53cd6c53
    10.25781/KAUST-I4JFX
    Scopus Count
    Collections
    Biological and Environmental Sciences and Engineering (BESE) Division; Bioscience Program; Bioscience Program; Theses

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