Identification and Investigation of Novel ALMS1 Interaction Partners in Cilia-Related Processes

DSpace Repository

Show simple item record

dc.contributor.advisor Rapaport, Doron (Prof. Dr.)
dc.contributor.author Wörz, Franziska Julia
dc.date.accessioned 2023-11-24T12:28:35Z
dc.date.available 2023-11-24T12:28:35Z
dc.date.issued 2023-11-24
dc.identifier.uri http://hdl.handle.net/10900/148069
dc.identifier.uri http://nbn-resolving.de/urn:nbn:de:bsz:21-dspace-1480694 de_DE
dc.identifier.uri http://dx.doi.org/10.15496/publikation-89409
dc.description.abstract Cilia, small antennae, can be found on nearly every cell in the human body and are important for human health and vision. Defects in cilia can lead to a variety of diseases, such as the Alström Syndrome (ALMS, OMIM #203800). This rare autosomal-recessive disorder occurs in 1 to 9 out of 1 million individuals. More than 300 mutations in the ALMS1 gene are known to cause a broad range of clinical defects most notably retinal dystrophy, Type 2 Diabetes, and truncal obesity. The ALMS1 gene codes for a ~0.5 MDa protein, that localizes to the centrosome and the basal body of cilia. So far, the molecular function of ALMS1 and its influence on the function of cilia is still elusive. Based on current literature and own data, I hypothesize that ALMS1 interacts with basal body proteins to enable proper cilia morphology and transport to the cilium. To address this hypothesis phenotypical and protein complex analyses were conducted. CRISPR/Cas9 mediated ALMS1 KO in hTERT-RPE1 cells was used to identify and decipher ALMS1 function in cilia biology. ALMS1-deficient cells presented shorter cilia and an impaired cellular proliferation compared to control cells. In addition, centrosomal proteins, such as γ-tubulin and CEP250, were investigated. While γ-tubulin remains unchanged, suggesting a function upstream of ALMS1, CEP250 localization was mildly but significantly reduced upon ALMS1 loss. Furthermore, cilia and cilia-related markers were examined, such as RPGR, which exhibits a compressed appearance, while others, such as IFT-88, were unchanged compared to the control. Further investigation is needed to understand the functional relevance of the observed phenotypic changes in the context of cilia biology as well as disease-related mechanisms. Furthermore, CRISPR/Cas9 mediated endogenously tagged ALMS1 HEK293T cells were used for protein complex analysis to further elucidate ALMS1 function. Investigation of protein-protein interaction revealed, that ALMS1 interacts among others with centrosomal and microtubule-associated proteins. Notably, the centrosomal protein 70 kDa (CEP70) was identified for the first time and appeared as a highly abundant ALMS1 interaction partner. Additional investigations, including deletion analysis, identified ALMS1 associating with the TPR domain and CT end of CEP70. By silencing CEP70 in hTERT-RPE1 cells a reduced ALMS1 localization at the basal body was observed, indicating CEP70 upstream of ALMS1. Nevertheless, further research is necessary to understand ALMS1 and CEP70 function in cilia-related processes, and disease-related mechanisms. en
dc.language.iso en de_DE
dc.publisher Universität Tübingen de_DE
dc.rights ubt-podno de_DE
dc.rights.uri http://tobias-lib.uni-tuebingen.de/doku/lic_ohne_pod.php?la=de de_DE
dc.rights.uri http://tobias-lib.uni-tuebingen.de/doku/lic_ohne_pod.php?la=en en
dc.subject.ddc 500 de_DE
dc.subject.other Alström syndrome en
dc.subject.other ALMS en
dc.subject.other ALMS1 en
dc.subject.other basal body en
dc.subject.other cilia en
dc.subject.other ciliopathy en
dc.subject.other protein complex analysis en
dc.subject.other ALMS1 protein network en
dc.title Identification and Investigation of Novel ALMS1 Interaction Partners in Cilia-Related Processes en
dc.type PhDThesis de_DE
dcterms.dateAccepted 2023-11-09
utue.publikation.fachbereich Biochemie de_DE
utue.publikation.fakultaet 7 Mathematisch-Naturwissenschaftliche Fakultät de_DE
utue.publikation.noppn yes de_DE

Dateien:

This item appears in the following Collection(s)

Show simple item record