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Stage-specific binding profile of cohesin in resting and switch-activated B lymphocytes

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  1. Tez No: 402648
  2. Yazar: GAMZE GÜNAL
  3. Danışmanlar: PROF. DR. ROLF JESSBERGER
  4. Tez Türü: Doktora
  5. Konular: Biyomühendislik, Bioengineering
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 2013
  8. Dil: İngilizce
  9. Üniversite: Technische Universität Dresden
  10. Enstitü: Yurtdışı Enstitü
  11. Ana Bilim Dalı: Belirtilmemiş.
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: 100

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Özet (Çeviri)

Cohesin is required for cell proliferation as it facilitates sister chromatid cohesion in dividing cells. Other regulatory functions of cohesin involve DNA repair, DNA recombination and transcriptional regulation. Cohesin also modulates antigen receptor gene rearrangements by facilitating higher-order chromatin interactions through various chromatin-associated factors. The surface expression of secondary immunoglobulin isotypes in B cells require the activation of essential transcription programs and class switch recombination by where the donor and acceptor S regions at the Igh locus are juxtaposed and the intervening DNA is irreversibly excised. This work aimed at elucidating the potential role of cohesin in class switch recombination and mature B cell specific gene expression, and regulation of chromatin at the Igh locus via the analysis of stage-specific binding profile of cohesin to chromatin in resting and switch-activated B lymphocytes. The first part of this study describes the significant, stage-dependent changes in cohesin binding to the B cell genome as well as its specific interactions during CSR- and at B-cell-related gene loci. In addition, the cohesin binding profile was compared to publicly available ChIP-seq profiles of the transcription regulator CTCF, and the transcription factor EBF1 in pro- and/or mature B cells. Cohesin and CTCF colocalize to the Igh locus in a stage-specific manner. EBF1, CTCF and cohesin are all found at the 3 Igh regulatory region. The finding of these potential regulatory sites will provide a concrete basis for the identification of new regulatory chromatin interactions. Moreover, cohesin is shown to be underrepresented at switch regions. Due to the lack of cohesin at S regions, facilitation of S/S synapsis by cohesin is unlikely. Cohesin underrepresentation at S regions suggest that S regions possess a cohesin-free structure to facilitate accessibility for AID and switch recombination machinery. The second part of this study aimed at establishing a mouse model for cohesin deficiency to study the potential role of cohesin in cells that undergo class switching. Cre transduced Smc3fl=fl B cells show reduced numbers of IgG1+ cells in vitro, due to increased apoptosis following SMC3-knockdown. A cohesin hypomorph phenotype could not be accomplished in animals with heterozygous mutations, since there was no difference in Smc3 mRNA and protein levels. Mice carrying the homozygous mutant were not viable. In conclusion, cohesin binds differentially to B-cell specific gene loci and to the Igh locus upon induction for class switching. These data suggest pathways by which gene expression is controlled, during the transition of IgM+ B cells into secondary Ig-secreting plasma cells, through the regulation of chromatin by cohesin.

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