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Identifier 000388606
Title Distinct roles of the Special AT-rich sequence-Binding protein-1 during T-cell development
Alternative Title Διακριτοί ρόλοι της SATB1 πρωτείνης κατά την ανάπτυξη των Τ-κυττάρων
Author Παναγόπουλος, Γεώργιος Π.
Thesis advisor Σπηλιανάκης, Χαράλαμπος
Abstract Special AT-rich sequence-binding protein 1 (SATB1) is a nuclear matrix protein with a central role in higher order chromatin organization and gene regulation. Elevated expression and/or mutation of SATB1 are associated with progression and poor prognosis in a plethora of malignancies. Although highly expressed in T cells, the molecular mechanism of SATB1 function in the thymus has been elusive. We identified protein interaction partners of SATB1 using a co-immunoprecipitation approach coupled with mass spectrometry analysis, using primary murine thymocyte extracts and created a comprehensive protein network map of SATB1 interactions in T cells. Among others, SATB1 interacts with several known transcription factors and kinases involved in long range chromatin interactions (YY1), thymic development, differentiation of naive CD4+ precursorcells (Mi-2β) and in the establishment of T cell tolerance (AIRE). Moreover, several members of specific histone modification complexes involved in gene regulation (COMPASS and NSL complexes) were also found to co-precipitate with SATB1 protein, suggesting an active role in the establishment/maintenance of T cell chromatin structure and gene regulation. Moreover, we optimized the protocol for the chromatin immunoprecipitation of the SATB1 protein in naive CD4+ T cells. Both the precipitated and the input DNA samples were sequenced in an Ion ProtonTM sequencer. We established a pipeline for downstream bioinformatics analysis and quality tests of next generation sequencing data for nuclear matrix proteins. A combination of the protein interactors of SATB1 and its binding sites on chromatin, could shed more light on the mechanism underlying the function of SATB1 in T cell development, chromatin conformation and gene regulation via long-range chromatin interactions.
Language English
Subject T-cells
Πρωτεϊνη
Τ-κύτταρα
Issue date 2014-11-21
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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