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.
| Identifer | oai:union.ndltd.org:kaust.edu.sa/oai:repository.kaust.edu.sa:10754/623470 |
| Date | 05 1900 |
| Creators | Al-Gashgari, Bothayna |
| Contributors | Fischle, Wolfgang, Biological and Environmental Sciences and Engineering (BESE) Division, Merzaban, Jasmeen, Mahfouz, Magdy M. |
| Source Sets | King Abdullah University of Science and Technology |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Rights | 2018-05-10, 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. |
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