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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 Κλαουράκης, Κωνσταντίνος
Thesis advisor Σπηλιανάκης, Χαράλαμπος
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.
Language Greek
Issue date 2016-07-22
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Graduate theses
  Type of Work--Graduate theses
Permanent Link https://elocus.lib.uoc.gr//dlib/8/c/b/metadata-dlib-1473692221-516090-4082.tkl Bookmark and Share
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