| dc.contributor.advisor | Wienands, Jürgen Prof. Dr. | |
| dc.contributor.author | Bhatt, Arshiya | |
| dc.date.accessioned | 2020-06-18T12:47:35Z | |
| dc.date.available | 2020-09-15T22:50:03Z | |
| dc.date.issued | 2020-06-18 | |
| dc.identifier.uri | http://hdl.handle.net/21.11130/00-1735-0000-0005-13DF-B | |
| dc.identifier.uri | http://dx.doi.org/10.53846/goediss-8045 | |
| dc.language.iso | eng | de |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
| dc.subject.ddc | 610 | de |
| dc.title | Unraveling details of CIN85/CD2AP assistance to SLP65-mediated B cell activation | de |
| dc.type | doctoralThesis | de |
| dc.contributor.referee | Wienands, Jürgen Prof. Dr. | |
| dc.date.examination | 2019-09-17 | |
| dc.description.abstracteng | The hallmark of adaptive immunity is the production of specific antibodies. This process is initiated upon the ligation of the B cell antigen receptor (BCR) which activates multiple intracellular signaling cascades through the phosphorylation of the B cell ‘master regulator’: SLP65. Our lab found that in order to function, SLP65 interacts constitutively – in a stimulation independent manner, with molecules of the CIN85/CD2AP-family of adaptor proteins, harbouring multiple protein interaction domains. Mouse models with B-cell specific CIN85 deletion and patients with a germline deletion in CIN85 encoding gene have shown that CIN85 is mandatory for B cell activation and the subsequent B cell responses. The molecular basis of the mechanism driving this, however, needed further elucidation. The primary cells pose a limitation to the scope of genetic and mechanistic studies due to the scarce availability of the reserve. I addressed these aspects through the generation of CIN85-deficient B cells from the established B cell lines, followed by subsequent deletion of CD2AP. The cellular system thus created was used to conduct genetic and biochemical functional assays to assess how these proteins work. I observed that CIN85 significantly impacts the Ca2+ flux upon BCR-ligation. Moreover, CIN85 could additionally affect an upstream NF-κB pathway signaling element called PKCβ. I also used the abovementioned CIN85/CD2AP-double deficient cells to investigate further binding partners of SLP65 employing SILAC-based mass spectrometry to look for possible functional redundancies. In our collaboration with Prof. C. Griesinger’s group, we found that CIN85 and SLP65 undergo phase-separation into supramolecular-clusters, representing the more recent and exciting intracellular compartments of membrane-free, mini-organelles, that orchestrate signaling in B cells. Due to their capacity to interact with multiple signaling elements, droplet signalosomes could be the modus operandi for CIN85/CD2AP-family of adaptor proteins for scaffolding of relevant signaling proteins. | de |
| dc.contributor.coReferee | Schwappach, Blanche Prof. Dr. | |
| dc.subject.eng | BCR signaling; B cell activation; SLP65; CIN85; CD2AP | de |
| dc.identifier.urn | urn:nbn:de:gbv:7-21.11130/00-1735-0000-0005-13DF-B-2 | |
| dc.affiliation.institute | Göttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB) | de |
| dc.subject.gokfull | Biologie (PPN619462639) | de |
| dc.description.embargoed | 2020-09-15 | |
| dc.identifier.ppn | 1701099136 | |