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dc.contributor.advisor Schuh, Melina Dr.
dc.contributor.author So, Chun
dc.date.accessioned 2019-10-01T11:28:55Z
dc.date.available 2019-10-01T11:28:55Z
dc.date.issued 2019-10-01
dc.identifier.uri http://hdl.handle.net/21.11130/00-1735-0000-0003-C1C9-0
dc.language.iso eng de
dc.relation.uri http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc 570 de
dc.title Optimization and application of Trim-Away for studying a liquid-like spindle domain in mammalian oocytes de
dc.type doctoralThesis de
dc.contributor.referee Schuh, Melina Dr.
dc.date.examination 2019-08-19
dc.description.abstracteng In this thesis, I reported a membraneless structure that permeated a major region of the female meiotic spindle and formed droplet-like protrusions around the spindle poles. Proteins within this previously unknown structure were highly dynamic and could redistribute rapidly throughout the entire spindle region. I found that this unusual structure behaves similar to a liquid and forms by phase separation and hence, termed it the liquid-like meiotic spindle domain (LISD). Interestingly, the LISD was not only present in the spindle of mouse oocytes, but also cow, pig and sheep oocytes and is thus widely conserved among mammals. Similar structures were not observed in wildtype or centrosome-depleted somatic cells, suggesting that the LISD is likely exclusive to the specialized spindle in oocytes. To identify the key proteins driving LISD assembly, I further optimized Trim-Away, the technique for acute depletion of endogenous proteins in mammalian cells. By combining in vitro and in vivo assays, I identified Aurora A kinase and two LISD proteins, transforming acidic coiled-coil-containing protein 3 (TACC3) and clathrin heavy chain 17 (CHC17), as the proteins essential for LISD assembly. Disruption of the LISD via different means released microtubule regulatory factors that reside within this domain into the cytosol and caused severe spindle defects. Spindles were half of their original size and failed to segregate chromosomes properly. Microtubule growth rates were reduced, and their overall turnover was increased. Both kinetochore fibers as well as interpolar microtubules were strongly depleted. A TACC3 mutant that loses its ability to phase-separate failed to rescue the substantial loss of microtubules in TACC3-depleted oocytes. Together, these data suggest that the LISD is necessary for the efficient assembly of stable acentrosomal spindle in mammalian oocytes. de
dc.contributor.coReferee Jakobs, Stefan Prof. Dr.
dc.contributor.thirdReferee Enderlein, Jörg Prof. Dr.
dc.contributor.thirdReferee Behr, Rüdiger Prof. Dr.
dc.contributor.thirdReferee Dosch, Roland Dr.
dc.contributor.thirdReferee Zweckstetter, Markus Prof. Dr.
dc.subject.eng LISD de
dc.subject.eng Liquid-like meiotic spindle domain de
dc.subject.eng Trim-Away de
dc.subject.eng Mammalian oocytes de
dc.subject.eng Meiotic spindle de
dc.subject.eng Meiosis de
dc.subject.eng Phase separation de
dc.subject.eng Microtubule de
dc.identifier.urn urn:nbn:de:gbv:7-21.11130/00-1735-0000-0003-C1C9-0-0
dc.affiliation.institute Göttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB) de
dc.subject.gokfull Biologie (PPN619462639) de
dc.identifier.ppn 1678020842

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