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    Chromatin association of UHRF1 during the cell cycle

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    Name:
    MS Thesis_Bothayna_Spring 2017.pdf
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    64.21Mb
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    Description:
    MS Thesis_Bothayna_Spring 2017
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    Type
    Thesis
    Authors
    Al-Gashgari, Bothayna cc
    Advisors
    Fischle, Wolfgang cc
    Committee members
    Merzaban, Jasmeen cc
    Mahfouz, Magdy M. cc
    Program
    Bioscience
    KAUST Department
    Biological and Environmental Science and Engineering (BESE) Division
    Date
    2017-05
    Embargo End Date
    2018-05-10
    Permanent link to this record
    http://hdl.handle.net/10754/623470
    
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    Access Restrictions
    At the time of archiving, the student author of this thesis opted to temporarily restrict access to it. The full text of this thesis became available to the public after the expiration of the embargo on 2018-05-10.
    Abstract
    Ubiquitin-like with PHD and RING Finger domains 1 (UHRF1) is a nuclear protein that associates with chromatin. Regardless of the various functions of UHRF1 in the cell, one of its more important functions is its role in the maintenance of DNA methylation patterns by the recruitment of DNMT1. Studies on UHRF1 based on this function have revealed the importance of UHRF1 during the cell cycle. Moreover, based on different studies various factors were described to be involved in the regulation of UHRF1 with different functionalities that can control its binding affinity to different targets on chromatin. These factors are regulated differently in a cell cycle specific manner. In light of this, we propose that UHRF1 has different binding behaviors during the cell cycle in regard to its association with chromatin. In this project, we first analyzed the binding behavior of endogenous UHRF1 from different unsynchronized cell systems in pull-down assays with peptides and oligonucleotides. Moreover, to analyze UHRF1 binding behavior during the cell cycle, we used two different approaches. First we sorted Jurkat and HT1080 cells based on their cell cycle stage using FACS analysis. Additionally, we synchronized HeLa cells to different stages of the cell cycle by chemical treatments, and used extracts from cellsorting and cell synchronization experiments for pull-down assays. We observed that UHRF1 in different cell systems has different preferences in regard to its binding to H3 unmodified and H3K9me3. Moreover, we detected that UHRF1, in general, displays different patterns between different stages of cell cycle; however, we cannot draw a final model for UHRF1 binding pattern during cell cycle.
    Citation
    Al-Gashgari, B. (2017). Chromatin association of UHRF1 during the cell cycle. KAUST Research Repository. https://doi.org/10.25781/KAUST-8I7XQ
    DOI
    10.25781/KAUST-8I7XQ
    ae974a485f413a2113503eed53cd6c53
    10.25781/KAUST-8I7XQ
    Scopus Count
    Collections
    Biological and Environmental Science and Engineering (BESE) Division; Bioscience Program; MS Theses

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