Graduate theses
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Title |
Uncovering a novel interaction between SATB1 - genome organizer and SET 1A - histone H3K4 methyltransferase |
Alternative Title |
Μελέτη της αλληλεπίδρασης της SATB1- οργανωτή χρωματίνης, και της SET1A- μεθυλοτρανσφεράσης ιστονών |
Author
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Κλαουράκης, Κωνσταντίνος
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Thesis advisor
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Σπηλιανάκης, Χαράλαμπος
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Abstract |
In our lab we study the effects of chromatin dynamics and epigenetic modifications in
the development of immune-related cells.
Nuclear matrix is highly involved in the
chromatin architecture
as DNA loci, known as
matrix attachment regions (MARs),
bind to it with the help of various proteins and
form
loops. The nuclear protein special
AT-rich sequence-binding protein 1 (SATB1) is highly expressed in T lymphocytes
and has
an important function in the formation of chromatin loops and genome
organization,
as well as in the regulation of gene expression.
SATB1 has been shown
to bind MARs with a specific sequence
in which
the one strand consists of mixed
A,
T, C nucleotides
and have a
strong potential for
unpairing when subjected to
superhelical strain.
Moreover,
mutations of SATB1
lead to autoimmunity and
various
cancers.
On the other hand histone modifications are one of the most important
mechanisms of transcription regulation. Among
the histone modifications mono-, di-
and tri-methylation of lysine 4 of histone 3 (H3K4me1/2/3)
have been
connected with
positive regulation of gene expression. In both mice and humans
the histone
modifications H3K4me1/2/3 are mediated by the proteins SET1A/B and MLL1/2/3
which are the catalytic
domains
of the COMPASS and COMPASS-like complexes
respectively.
Previous work in our lab
has
utilized
immunoprecipitation
coupled to
mass spectrometry
analysis
(IP-MS)
in
murine
thymocytes to identify SATB1’s
protein interactome. These experiments indicated, among
other interactions, an
interaction between SATB1 and COMPASS, as well as COMPASS-like complexes.
Therefore, it was
hypothesized
that SATB1 interacts
and guides the COMPASS
complex to specific loci in order to methylate histone H3K4 and induce the
expression of specific genes.
This work
tested
the interaction of SATB1 with the
histone methyltransferase SET1A in
murine
thymocytes, first by double
immunofluorescence-sequential staining and then by co-immunoprecipitation coupled
to
Western blot (co-IP-WB). Moreover
in order to
identify changes of
the histone
methylation
pattern in the absence of SATB1
we performed
chromatin
immunoprecipitation
coupled with
next generation
sequencing
(ChIP-Seq)
experiments
for the histone modification H3K4me2 in C57BL/6
and
CD4. Cre-Satb1 fl/fl
thymocytes. Finally
I created
an online automated
workflow
in order to
analyze
raw
ChIP-Seq data.
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Language |
Greek |
Issue date |
2016-07-22 |
Collection
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School/Department--School of Sciences and Engineering--Department of Biology--Graduate theses
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Type of Work--Graduate theses
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Permanent Link |
https://elocus.lib.uoc.gr//dlib/8/c/b/metadata-dlib-1473692221-516090-4082.tkl
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Views |
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