Regulation of the activity of ubiquitin ligase HACE1

Toplak, Julia Jelena

prikaz prve stranice dokumenta Regulation of the activity of ubiquitin ligase HACE1

Embargo period 2025-09-21

master's thesis

Regulation of the activity of ubiquitin ligase HACE1

Zagreb: University of Zagreb, Faculty of Science, 2023. urn:nbn:hr:217:358497

Toplak, Julia Jelena

University of Zagreb
Faculty of Science
Department of Biology

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APA 6th Edition

Toplak, J. J. (2023). Regulation of the activity of ubiquitin ligase HACE1 (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:358497

MLA 8th Edition

Toplak, Julia Jelena. "Regulation of the activity of ubiquitin ligase HACE1." Master's thesis, University of Zagreb, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:217:358497

Chicago 17th Edition

Toplak, Julia Jelena. "Regulation of the activity of ubiquitin ligase HACE1." Master's thesis, University of Zagreb, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:217:358497

Harvard

Toplak, J. J. (2023). 'Regulation of the activity of ubiquitin ligase HACE1', Master's thesis, University of Zagreb, Faculty of Science, accessed 26 January 2025, https://urn.nsk.hr/urn:nbn:hr:217:358497

Vancouver

Toplak JJ. Regulation of the activity of ubiquitin ligase HACE1 [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2023 [cited 2025 January 26] Available at: https://urn.nsk.hr/urn:nbn:hr:217:358497

IEEE

J. J. Toplak, "Regulation of the activity of ubiquitin ligase HACE1", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2023. Available at: https://urn.nsk.hr/urn:nbn:hr:217:358497

Cite this item: https://urn.nsk.hr/urn:nbn:hr:217:358497

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Metadata

Title	Regulation of the activity of ubiquitin ligase HACE1
Title (croatian)	Regulacija aktivnosti ubikvitinske ligaze HACE1
Author	Julia Jelena Toplak
Mentor	Sonja Lorenz (mentor)
Mentor	Dunja Leljak-Levanić (komentor)
Committee member	Dunja Leljak-Levanić (predsjednik povjerenstva)
Committee member	Biljana Balen (član povjerenstva)
Committee member	Ivana Ivančić Baće (član povjerenstva)
Committee member	Petra Peharec Štefanić (član povjerenstva)
Granter	University of Zagreb Faculty of Science (Department of Biology) Zagreb
Defense date and country	2023-09-21, Croatia
Scientific / art field, discipline and subdiscipline	NATURAL SCIENCES Biology
Abstract	The HECT ubiquitin ligase HACE1 is involved in membrane dynamics regulation, mitophagy, redox homeostasis, and cytoskeleton organization. Despite HACE1 being a tumor suppressor and a significant factor in neural development, it remains biochemically poorly characterized, and its regulatory mechanisms are not known. This master’s thesis explores the impact of phosphorylation as a regulatory mechanism and its effect on HACE1 activity and dimerization. Phosphomimetic variants were generated and characterized using functional assays, mass photometry, and analytical size exclusion chromatography. The findings reveal that the phosphomimetic mutations within the N-terminal helix disrupted dimerization and activated HACE1 activity, providing insights into how the monomer could be stabilized in its active state. Furthermore, recombinantly purified kinases PAK1 and TBK1 were investigated as potential regulators of HACE1 activity, and the sites targeted by these kinases in HACE1 sites were identified through mass spectrometry.
Abstract (croatian)	Ubikvitinska HECT ligaza HACE1 sudjeluje u regulacije dinamike membrana, mitofagiji, redokshomeostazi i organizaciji citoskeleta. Unatoč tome što je ligaza HACE1 tumor-supresor i značajan faktor u razvoju živčanog sustava, nije detaljno biokemijski karakterizirana, a regulatorni mehanizmi nisu poznati. Ovaj diplomski rad istražuje utjecaj fosforilacije kao regulatornog mehanizma na aktivnost i dimerizaciju HACE1. Pripremljene su fosfomimetičke varijante i karakterizirane korištenjem funkcionalnih eseja, masene fotometrije i analitičke kromatografije temeljene na isključivanju. Rezultati otkrivaju da fosfomimetičke varijante unutar N-terminalnog heliksa narušavaju dimerizaciju i aktiviraju aktivnost ligaze HACE1, pružajući uvid u to kako se monomer može stabilizirati u aktivnom stanju. Nadalje, rekombinantno pročišćene kinaze PAK1 i TBK1 istražene su kao potencijalni regulatori. Mjesta u ligazi HACE1 koje te kinaze ciljaju identificirana su putem masene spektrometrije.
Keywords
Keywords (croatian)
Language	english
URN:NBN	urn:nbn:hr:217:358497
Study programme	Title: Molecular Biology Study programme type: university Study level: graduate Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resource	Text
File origin	Born digital
Access conditions	Embargoed access Embargo expiration date: 2025-09-21
Terms of use
Repository	Repository of the Faculty of Science
Created on	2023-10-25 10:22:08