| dc.contributor.advisor | Rehling, Peter Prof. Dr. | |
| dc.contributor.author | Müller, Tobias | |
| dc.date.accessioned | 2019-12-17T11:44:39Z | |
| dc.date.available | 2019-12-17T11:44:39Z | |
| dc.date.issued | 2019-12-17 | |
| dc.identifier.uri | http://hdl.handle.net/21.11130/00-1735-0000-0005-12D2-9 | |
| dc.identifier.uri | http://dx.doi.org/10.53846/goediss-7776 | |
| dc.language.iso | eng | de |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
| dc.subject.ddc | 610 | |
| dc.title | Role of MICOS for mitochondrial morphology and function | de |
| dc.type | doctoralThesis | de |
| dc.contributor.referee | Jahn, Olaf Dr. | |
| dc.date.examination | 2019-07-01 | |
| dc.description.abstracteng | Mitochondria are highly dynamic organelles with a distinct morphological membrane
ultrastructure. The mayor protein complex responsible for the formation and maintenance
of the inner mitochondrial membrane is the mitochondrial contact site and cristae
organizing system (MICOS). This complex consists of 6 constituents in Saccharomyces
cerevisiae and 7 in the mammalian system, with Mic60 and Mic10 being the core components.
MICOS has been predominantly investigated in S. cerevisiae and studies in
human cells have focussed mainly on MIC60.
In this study, a CRISPR/CAS9 mediated MIC10 knock-out cell-line was generated
and further characterized. Herby MIC10 could be identified to be essential for forming
and maintaining proper mitochondrial morphology in mammalian cells. However, the
loss of inner membrane ultrastructure did not have an impact on mitochondrial function
and health.
To better understand MICOS function in human, two different proximity biotinylation
approaches were undertaken and compared to determine novel interaction partners. The
first approach involved using an enhanced ascorbate C peroxidase APEX fused to MIC10
and the second approach made use of a promiscuous biotin ligase BioID2. The usage of
the BioID2 enzyme proved to be more suitable since it already produced a distinct set
of mutual interaction partners together with already known interaction partners. On
the other hand, the active labelling reagent using the APEX enzyme proved to be more
reactive than anticipated and would need a more thorough control system to identify
background labelling.
Recent findings reported a connection between MICOS via Mic60 and protein translocation
through the TOM complex and the MIA-pathway (von der Malsburg et al., 2011).
In this study, a spatial connection between MICOS and the TIM23 complex mediated
via Mic60 could be found. Performing import studies in two different yeast strains with
impaired inner membrane ultrastructure mic10Δ and atp20Δ revealed, that independent
from MICOS, proper inner mitochondrial membrane morphology is essential for efficient
precursor protein translocation via the TIM23-complex. In contrast to the mammalian
system, carrier import via the TIM22 complex was verified to be independent of morphological
alterations in yeast, thus further confirming the immense evolutionary divergence
between the human and yeast TIM22 carrier translocase. | de |
| dc.contributor.coReferee | Meinecke, Michael Prof. Dr. | |
| dc.subject.eng | MICOS | de |
| dc.subject.eng | Translocase | de |
| dc.identifier.urn | urn:nbn:de:gbv:7-21.11130/00-1735-0000-0005-12D2-9-1 | |
| dc.affiliation.institute | Medizinische Fakultät | |
| dc.subject.gokfull | Biochemie / Physiologische Chemie / Pathobiochemie - Allgemein- und Gesamtdarstellungen (PPN619875313) | de |
| dc.identifier.ppn | 1685861091 | |