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master's thesis
Karakterizacija proteinskih kompleksa proteina SSB
Zagreb: University of Zagreb, Faculty of Science, 2019. urn:nbn:hr:217:603412
Kostelac, Anja
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Kostelac, A. (2019). Karakterizacija proteinskih kompleksa proteina SSB (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:603412

Kostelac, Anja. "Karakterizacija proteinskih kompleksa proteina SSB." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:603412

Kostelac, Anja. "Karakterizacija proteinskih kompleksa proteina SSB." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:603412

Kostelac, A. (2019). 'Karakterizacija proteinskih kompleksa proteina SSB', Master's thesis, University of Zagreb, Faculty of Science, accessed 21 May 2024, https://urn.nsk.hr/urn:nbn:hr:217:603412

Kostelac A. Karakterizacija proteinskih kompleksa proteina SSB [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2019 [cited 2024 May 21] Available at: https://urn.nsk.hr/urn:nbn:hr:217:603412

A. Kostelac, "Karakterizacija proteinskih kompleksa proteina SSB", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2019. Available at: https://urn.nsk.hr/urn:nbn:hr:217:603412

Metadata
TitleKarakterizacija proteinskih kompleksa proteina SSB
Title (english)Characterisation of SSB protein complexes
AuthorAnja Kostelac
MentorDušica Vujaklija (mentor)
MentorIvana Ivančić Baće (mentor)
Committee memberIvana Ivančić Baće (predsjednik povjerenstva)
Committee memberSunčica Bosak (član povjerenstva)
Committee memberAna Previšić (član povjerenstva)
Committee memberZlatko Liber (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2019-02-18, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Streptomiceti predstavljaju najbrojniji rod unutar koljena Actinobacteria. Bakterija Streptomyces coelicolor A3(2) je modelni organizam za proučavanje procesa stanične diferencijacije i proizvodnje antibiotika kod bakterija. Genom ove bakterije ima dva gene, ssbA i ssbB za proteine koji vežu jednolančanu molekulu DNA (SSB), a sudjeluju u različitim procesima metabolizma DNA. Također ima i dva gena, hupA i hupS, koji kodiraju za proteine HU. Ovi proteini su združeni s nukleoidom i važni su za očuvanje strukture i integriteta DNA unutar stanice. Protein HU je potencijalni proteinski partner proteina SsbA. U ovom je radu, proteini SsbA i HupS prekomjerno su eksprimirani u heterolognom domaćinu, bakteriji Escherichia coli u svrhu pročišćavanja veće količina proteina. Za potrebe ispitivanja kompleksa SSB:HU ispitane su interakcije protein-DNA proteina SsbA i SsbB s jednolančanom DNA. Rezultati su pokazali da protein SsbB ima prosječno dva puta veći afinitet prema jlDNA nego protein SsbA, te da nastajanje kompleksa SsbB:jlDNA ne ovisi značajnije o prisutnosti soli u odnosu na kompleks SsbA:jlDNA. Pokazano je da je nastajanje kompleksa SsbA:HU spontana reakcija s visokom konstantom disocijacije i da je vođena hidrofobnim interakcijama. Interakcija između proteina SsbA i HU se ostvaruje bez prisustva jlDNA, dok se konformacijska promjena uočava samo na proteinu HU.
Abstract (english)
Streptomyes are the largest genus of Actinobacteria. Streptomyces coelicolor A3(2) is a model organism for the study of bacterial differentiation and antibiotic production. This bacterium has two genes, ssbA and ssbB, encoding single-stranded DNA binding proteins (SSBs) which participate in various processes of DNA metabolism. It also possesses two genes, hupA and hupS, encoding nucleoid-associated HU proteins which are involved in maintaining DNA integrity in the cell. In previous study we identified HU protein as a potential SSB protein partner. In this study, SsbA and HupS proteins were overexpressed in heteroulogous host, Escherichia coli and a large amount of proteins was purified for further analyses. In order to characterise complex SsbA:HU, protein-DNA interactions were analysed between proteins SsbA and SsbB and single-stranded DNA (ssDNA). The results showed protein SsbB has two-fold increased affinity with ssDNA compared to protein SsbA and that formation of SsbB:ssDNA is less salt dependent compared to SsbA:ssDNA. This study showed that formation of SsbA:HU complex is a spontaneous reaction with high dissociation constant, which is driven by hydrophobic interactions. Protein-protein interactions between SsbA and HU are formed without the presence of ssDNA, whereas conformational changes were observed only for HU protein.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:603412
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2021-02-18
Terms of useLicence In copyright icon
Created on2019-04-02 08:28:21