Karakterizacija Ankle1, nove ljudske endonukleaze koja sadrži LEM-domenu

master's thesis

• Title (english): Characterization of Ankle1 , a novel human LEM-domain containing endonuclease
• Author: Livija Zlopaša
• Mentor: Zdravko Lorković (mentor)
• Committee member: Zdravko Lorković (predsjednik povjerenstva)
• Committee member: Željka Vidaković-Cifrek (član povjerenstva)
• Committee member: Gordana Lacković-Venturin (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2011-09-22, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

LEM-proteini čine skupinu jezgrinih proteina sa zajedničkim strukturnim motivom dugim oko 40 aminokiselina nazvanim LEM-domena (ime je izvedeno iz imena proteina unutarnje jezgrine membrane: LAP2, emerin i MAN1). LEM-domena svih dosad analiziranih proteina posreduje u direktnom vezanju za BAF (barrier-to-autointegration factor). BAF je evolucijski visoko očuvani protein koji veže dvolančanu DNA, proteine jezgrine lamine, histone, te različite transkripcijske faktore. LEMproteini i BAF su otkriveni u višestaničnim organizmima, a nisu prisutni u jednostaničnim organizmima i biljkama. Ankle1 je novi i neobični protein koji sadrži LEM-domenu te, za razliku od većine ostalih LEMproteina, nije dio jezgrine membrane već se nalazi u nukleoplazmi. Predviđanje organizacije domena pokazuje da je Ankle1 očuvan tijekom evolucije od Caenorhabditis elegans do Homo sapiens. Ankle1 sadrži ankirinska ponavljanja, za koje se vjeruje da posreduju prilikom stvaranja interakcija među proteinima, LEM-domenu i GIY-YIG domenu, koja je opisana kao domena sa nukleaznom aktivnošću u skupini proteina nazvanih GIY-YIG endonukleaze. Predviđanja in silico otkrila su nukleotidne sljedove koji posreduju transport u i iz jezgre, koji se nalaze na očuvanim pozicijama unutar Ankle1 proteina, te koji nagoviještaju da se Ankle1 premješta između jezgre i citoplazme. Analiza uzorka ekspresije pokazala je da se Ankle1 izražava primarno u hematopoetskim tkivima, nagovještavajući moguću funkciju Ankle1 proteina tijekom razvoja i diferencijacije hematopoetskih staničnih linija.

Abstract (english)

LEM domain proteins are a group of nuclear proteins that share a common structural motif of approximately 40 amino acids, known as LEM domain (the name is derived from inner nuclear membrane proteins LAP2, emerin and MAN1). The LEM domain of all analysed proteins mediates association through direct binding to barrier-to-autointegration factor (BAF). BAF is a highly conserved protein that binds to dsDNA, nuclear lamina proteins, histones and various transcription factors. LEM domain proteins and BAF have been identified in multicellular organisms but are absent in unicellular eukaryotes and plants. Ankle1 is a novel unusual LEM domain protein which is, unlike most of the other LEM domain proteins, not an integral membrane protein, but localizes to the nucleoplasm. Prediction of its domain organization shows that Ankle1 is conserved throughout evolution from Caenorhabditis elegans to Homo sapiens. It contains ankyrin repeats, a LEM domain and a GIY-YIG domain, which has previously been described as a nuclease domain in a protein family called GIY-YIG endonucleases. In silico predictions revealed nuclear export sequences (NES) and nuclear localization sequences (NLS) at conserved positions within the Ankle1 protein, indicating that the protein shuttles between nucleus and cytoplasm. Expression pattern analysis showed that Ankle1 is expressed primarily in haematopoietic tissues and cell lines derived thereof, indicating a potential function during the development and differentiation of haematopoietic cells lineages.

• Keywords: Ankle1; LEM-proteini; NES; NLS; GIY-YIG nukleazna domena
• Keywords (english): Ankle1; LEM proteins; NES; NLS. GIY-YIG nuclease domain
• Language: croatian
• URN:NBN: urn:nbn:hr:217:625497
• 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: Access restricted to students and staff of home institution
• Created on: 2017-05-03 11:58:54