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doctoral thesis
Molekularna analiza elemenata interakcije FNR-TROL
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:383022
Rac, Anja
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Rac, A. (2021). Molekularna analiza elemenata interakcije FNR-TROL (Doctoral thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:383022

Rac, Anja. "Molekularna analiza elemenata interakcije FNR-TROL." Doctoral thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:383022

Rac, Anja. "Molekularna analiza elemenata interakcije FNR-TROL." Doctoral thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:383022

Rac, A. (2021). 'Molekularna analiza elemenata interakcije FNR-TROL', Doctoral thesis, University of Zagreb, Faculty of Science, accessed 03 January 2025, https://urn.nsk.hr/urn:nbn:hr:217:383022

Rac A. Molekularna analiza elemenata interakcije FNR-TROL [Doctoral thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2025 January 03] Available at: https://urn.nsk.hr/urn:nbn:hr:217:383022

A. Rac, "Molekularna analiza elemenata interakcije FNR-TROL", Doctoral thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:383022

Metadata
TitleMolekularna analiza elemenata interakcije FNR-TROL
Title (english)Molecular analysis of the FNR-TROL interaction elements
AuthorAnja Rac
MentorHrvoje Fulgosi (mentor)
Committee memberŽeljka Vidaković Cifrek (predsjednik povjerenstva)
Committee memberĐurđica Ugarković (član povjerenstva)
Committee memberLea Vojta (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Universal decimal classification
(UDC)		577 - Biochemistry. Molecular biology. Biophysics
Abstract
U ovom doktorskom radu istraţena je uloga interakcije FNR-TROL u transportnom lancu elektrona na tilakoidima u kloroplastu. Feredoksin-NADP+ oksidoreduktaza (FNR) je protein koji katalizira posljednji korak u transportu elektrona- prijenos elektrona s reduciranog feredoksina na NADP+ pri čemu nastaje NADPH. Nalazi se u obliku topljivih izoformi ili vezan za tilakoidnu membranu čineći proteinske komplekse s proteinima Tic62 i TROL. U svrhu istraţivanja utjecaja kompleksa s proteinom TROL korištene su mutantne biljke Arabidopsis thaliana koje nisu eksprimirale protein TROL, ali su sadrţavale FNR1 izoformu iz C4 biljke kukuruza, koja je pokazala pojačan afinitet za vezanje u komplekse s proteinom TROL. Te su biljke kriţane s mutantnim biljkama koje su sadrţavale TROL s izmijenjenim ili uklonjenim aminokiselinama rodanazne domene za koju se smatra da ima vaţnu ulogu u signalizaciji otpuštanja i vezanja proteina FNR čime regulira tok elektrona u transportnom lancu. Kvantitativna i kvalitativna analiza jačine interakcija proteina TROL s izoformama proteina FNR1 iz kukuruza i graška provedena je sustavom dvaju hibrida kvasaca te praćena testovima aktivnosti X-alpha i beta galaktozidaze. Provjera interakcije proteina u kvascima pokazala je snaţnije interakcije proteina TROL i Tic62 s FNR1 izoformom kukuruza u odnosu na one s FNR1 izoformom graška. Utjecaj izmijenjenih domena proteina TROL i interakcija s kukuruznim proteinom FNR1 ispitan je Westernskom metodom otiska, odreĎivanjem koncentracije pigmenata te analizom anatomske strukture i ultrastrukture listova.Pokazano je da je kukuruzni FNR1 vezan na tilakoidima samo kod kriţanca kod kojeg nije bilo promjene u domeni RHO što upućuje na neospornu vaţnost lumenalne domene RHO u transmembranskoj signalizaciji vezanja i otpuštanja proteina FNR na stromalnoj strani tilakoida. Daljnja istraţivanja i rasvjetljavanje puta prijenosa signala povezanog s domenom RHO mogao bi doprinijeti razumijevanju alternativne raspodjele elektrona u fotosintezi vaskularnih biljaka.
Abstract (english)
In this thesis, the role of the FNR-TROL interaction in electron transport chain on thylakoid membranes was investigated. Ferredoxin-NADP+ oxidoreductase (FNR) catalyses the last step in electron transport chain from reduced ferredoxin to NADP+ forming NADPH. FNR isoforms can be found as soluble, or form protein complexes with proteins Tic62 or TROL on thylakoid membranes. To investigate complex with TROL protein A. thaliana trol plants which do not express protein TROL but contain C4 maize FNR1, which showed enhanced binding affinity to TROL, were crossed with mutant plants in which RHO domain of TROL protein has been modified. RHO domain is considered as important in FNR binding and release signaling, therefore modifications in mutant plants included changed or deleted aminoacids. Qualitative and quantitative analysis of binding maize and pea FNR1 into complexes with TROL was investigated with Yeast two hybrid screening following X-alpha and beta galactosidase activity assays. Assays have shown stronger interactions between TROL and Tic62 with maize FNR1 in relation to interactions with pea FNR1. Effect of changed RHO domain and interaction with maize FNR1 were investigated with Western blot coupled with ECL, by measuring pigment concentration, and by comparing anatomical leaf structure and ultrastructure. Maize FNR1 was found on thylakoids only in mutant line in which RHO domain wasn't modified. This result implies an important role of RHO domain on transmembrane signaling from lumenal side to stromal side of thyakoids and it's effect on FNR-TROL interactions. Future investigations on lumenal mechanism that triggers transmembrane signal via RHO domain might give better understanding of this signal pathways in alternative electron transport chain.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:383022
Promotion2021
Study programmeTitle: Biology
Study programme type: university
Study level: postgraduate
Academic / professional title: doktor/doktorica znanosti, područje prirodnih znanosti, polje biologija (doktor/doktorica znanosti, područje prirodnih znanosti, polje biologija)
Type of resourceText
File originBorn digital
Access conditionsOpen access
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Created on2021-03-16 12:31:31