Browsing by Author "Bulatović, Luka"

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    Description of the cyclin Clb5 docking motif in Far1
    (Tartu Ülikool, 2019) Bulatović, Luka; Faustova, Ilona, supervisor; Örd, Mihkel, supervisor
    In English: Cell cycle is a timely ordered, multistep event during which a cell’s chromosomal DNA is first replicated and later divided into two identical daughter cells. Cyclin-dependent kinases (Cdks) regulate the events of the cell cycle through phosphorylation of different substrates. Cdks are activated by binding to an activating partner – a cyclin. Cyclins are also substrate specific, i.e. they bind to a substrate and promote its phosphorylation by the Cdk complex. In this work, we mapped a newly discovered Clb5 cyclin docking motif present in the N – terminal region of a Cdk inhibitor protein, Far1. We found that mutations in the motif dras-tically delay phosphorylation-dependent degradation rates of Far1. In addition to this, we showed that Far1 docking region functions as an independent linear docking motif. Interest-ingly, our data also suggests that the motif does not bind to the hydrophobic patch of Clb5, suggesting that there might be some other, not yet discovered, binding pocket on the cyclin. Eesti keeles: Rakutsükkel on mitmeosaline järjestikkune protsess, mille käigus raku DNA esmalt paljun-datakse ja seejärel jagatakse kahe tütarraku vahel. Tsükliinist sõltuvad kinaasid (Cdk) reg-uleerivad rakutsükli sündmusi erinevate substraatide fosforüleerimise kaudu. Cdk on ak-tiivne ainult kompleksis tsükliiniga, kusjuures tsükliinid suunavad kinaasi kindlaid valke fosforüleerima. Käesolevas töös leiti, et Cdk inhibiitorvalgu Far1 N-terminaalses alas paikneb tsükliini Clb5 seondumismotiiv. Selle motiivi muteerimine põhjustab suure hiline-mise Far1 fosforüleerimisest sõltuvas lagundamises rakutsükli käigus. Lisaks näidati, et Far1 seondumismotiiv toimib iseseisva lineaarse motiivina, mis toimib tsükliini Clb5 hüdrofoob-sest taskust sõltumatult, vihjates, et tsükliini pinnal on teisigi substraatide sidumistaskuid.