Homologna i heterologna ekspresija gena AtBPM1 u transgeničnim biljkama Arabidopsis thaliana i transformiranim stanicama Nicotiana tabacum

master's thesis

• Title (english): Homologous and heterologous expression of AtBPM1 in Arabidopsis thaliana plants and in transformed Nicotiana tabacum cells
• Author: Jelena Martinčić
• Mentor: Nataša Bauer (mentor)
• Committee member: Nataša Bauer (predsjednik povjerenstva)
• Committee member: Mirta Tkalec (član povjerenstva)
• Committee member: Mirjana Kalafatić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2012-02-03, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Proteini s proteinskim domenama BTB/POZ su identificirani u mnogih eukariota gdje su im definirane raznolike funkcije. Takvi proteini mogu sadržavati i dodatne domene, kao što je domena MATH. Domene MATH/BTB (BPM) proteinima biljaka Arabidopsis thaliana omogućuju različite proteinske interakcije. Pretpostavlja se da protein AtBPM1 sudjeluje u prepoznavanju supstrata i njegovoj ubikvitinaciji u sklopu ubikvitin ligaza E3 koje sadrže kulin 3 (CUL3). Točna funkcija proteina AtBPM1 nije definirana. Kako bi pridonijeli razumijevanju funkcije proteina AtBPM1, u ovom radu je istražena njegova lokalizacija u transgeničnim biljkama uročnjaka te u transformiranim stanicama duhana BY-2. Kotransformacijom stanica BY-2 metodom Biolistic® utvrđeno je da proteini AtBPM1 i ZmCUL3 kolokaliziraju u citoplazmi, a manjim dijelom u jezgri. Protein AtBPM1 lokalizira najvećim dijelom u jezgrici gdje mu je funkcija najvjerojatnije neovisna o CUL3. Nije utvrđeno postojanje stabilnih homodimera i heterodimera proteina AtBPM1, kao ni ubikvitinacija navedenog proteina. Tretman inhibitorom proteasoma MG132 rezultirao je nakupljanjem proteina AtBPM1 i ZmCUL3, što ukazuje na razgradnju navedenih proteina proteasomima.

Abstract (english)

BTB/POZ proteins are identified in many eukaryotes in which they obtain different functions. Such proteins can have some additional domains, like MATH domain. Those domains enable Arabidopsis thaliana MATH/BTB (BPM) proteins to have different protein-protein interactions. It is assumed that AtBPM1 protein participates in substrate recognition and ubiquitination in CUL3 E3 ubiquitin ligases. The exact function of AtBPM1 protein is not defined. In order to contribute to the knowledge of AtBPM1 protein function, the aim of this study was to investigate its localization in transgenic A. thaliana plants and in transformed tobacco BY-2 cells. Due to Biolistic® transformation of BY-2 cells, it is determined that AtBPM1 and ZmCUL3 proteins co-localize in cytoplasm, and partly in nucleus. AtBPM1 protein localizes to a large extent in nucleolus, where its function is probably independent of CUL3. Stable homodimers or heterodimers and ubiquitinated AtBPM1 protein is not found. Treatment with proteasome inhibitor MG132 lead to accumulation of AtBPM1 and ZmCUL3, what indicates on involvement of proteasome in degradation of examined proteins.

• Keywords: proteinske domene MATH/BTB; transgenične biljke Arabidopsis thaliana; ubikvitinacija proteina; ubikvitin ligaze E3 koje sadrže CUL3
• Keywords (english): MATH/BTB protein domains; Arabidopsis thaliana transgenic plants; protein ubiquitination; CUL3 E3 ubiquitin ligases
• Language: croatian
• URN:NBN: urn:nbn:hr:217:012699
• 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: 2017-05-02 09:36:57