Optimization and application of Trim-Away for studying a liquid-like spindle domain in mammalian oocytes

So, Chun

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.identifier.uri	http://dx.doi.org/10.53846/goediss-7653
dc.language.iso	eng	de
dc.rights.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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Das Dokument erscheint in:

GGNB - Göttinger Graduiertenzentrum für Neurowissenschaften, Biophysik und molekulare Biowissenschaften [1115]
GGNB - Göttingen Graduate Center for Neurosciences, Biophysics and Molecular Biosciences

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