University of Zagreb. Faculty of Science. Department of Chemistry.
Abstract
Na početku mitoze mikrotubuli i pripadajući proteini formiraju diobeno vreteno, strukturu
koja omogućuje točnu raspodjelu kromosoma između dvije stanice kćeri. U ovom radu
razvijena je optogenetička metoda koja omogućuje brzu i reverzibilnu translokaciju proteina
iz diobenog vretena na mitohondrije korištenjem plavog svjetla. Metoda je primijenjena za
proučavanje proteina TACC3 i PRC1 koji sudjeluju u formiranju paralelnih, odnosno
antiparalelnih svežnjeva mikrotubula. Translokacija oba proteina događala se na vremenskim
skalama od nekoliko minuta. Mijenjanjem koncentracije proteina PRC1 na diobenom vretenu
bilo je moguće inducirati raspadanje i ponovno formiranje antiparalelnih svežnjeva
mikrotubula što je rezultiralo promjenom oblika diobenog vretena. Dakle, ova metoda
predstavlja novi moćni alat za proučavanje proteina diobenog vretena kontrolom njihove
lokalizacije u pojedinim fazama mitoze.At the onset of mitosis microtubules and microtubule associated proteins form a spindle,
which is responsible for proper segregation of chromosomes between two daughter cells. In
this thesis, I have developed optogenetic method for rapid and reversible translocation of
proteins from the mitotic spindle to mitochondria using blue light. This method was used for
studying TACC3 and PRC1, proteins which are required for formation of parallel and
antiparallel microtubule bundles, respectively. Translocation of both proteins occurred at time
scales of several minutes. By changing concentration of PRC1 in the mitotic spindle, I was
able to induce disassembly and formation of antiparallel microtubule bundles, which resulted
in changes in the spindle shape. Thus, this method represents powerful new tool for studying
spindle proteins by controlling their localization at specific stages of mitosis