izvor podataka: poirot

Naziv

MATH-BTB proteini kao regulatori transkripcije i RNA posredovane metilacije DNA u biljnom razvitku

MATH-BTB proteins as switches between transcription and RNA-directed DNA methylation during plant development

Opis projekta

Metilacija DNA jedan je od nekoliko epigenetičkih mehanizama kontrole ekspresije gena. U biljaka raznolikost epigenetičkih mehanizama ima, osim u regulaciji razvojnih procesa, važnu ulogu u fenotipskoj i fiziološkoj prilagodljivosti, te sposobnosti biljaka da prežive i razmnožavaju se u promjenjivom okolišu. Metilacija biljne DNA odvija se na sljedovima CG, CHG i asimetričnom CHH (H simbol za A, C ili T), a svaki od njih ima različite ''genetičke zahtjeve'' za očuvanje. Nesimetrična metilacija CHH ne može se održati i zahtijeva de novo metilaciju u svakom staničnom ciklusu. Proces kojim se to ostvaruje je metilacija DNA usmjerena s RNA (RdDM). Dvije RNA polimeraze specifične za biljke, Pol IV i Pol V sintetiziraju male interferirajuće RNA, odnosno dugačke nekodirajuće molekule RNA, važne za RNA-DNA prepoznavanje. Kompleks DDR predstavlja proteinski kompleks za koji se smatra da je odgovoran za dovođenje enzima Pol V na mjesto regulacije gena, no mehanizam specifičnosti nije jasan. Naši preliminarni rezultati ukazuju da proteini MATH-BTB iz uročnjaka stupaju u interakciju s kompleksom DDR, te tako mogu posredovati u pozicioniranju sustava RdDM. Najnovija otkrića ukazuju da su supstrati proteina BPM transkripcijski faktori. Proteini s ovakvim interakcijskim mogućnostima imaju potencijal održavanja ravnoteže između metilacije (posredovane Pol V) i transkripcije (posredovane Pol II). Glavni cilj projekta je rasvjetljavanje funkcionalne uloge proteina BPM u regulaciji transkripcije i RdDM-a kao i moguće uloge u biljnoj embriogenezi i prilagodbi na stresne uvjete. Posebna pozornost biti će dana identifikaciji gena čija je transkripcija ili RdDM metilacija uravnotežena proteinom BPM1 (kromatinska imunoprecipitacija-ChIP, ChIP-sekvenciranje). Nadalje, metilacijski uzorak (bisulfitno sekvenciranje) odabranih gena će biti povezan s njihovom ekspresijom (Q-PCR), te s embriogenezom i odgovorom biljaka na stres u linijama s promijenjenim sustavom RdDM ili ekspresijom gena BPM.

DNA methylation is one of several epigenetic mechanisms controlling gene expression. In plants, the diversity of epigenetic mechanisms has, in addition to the regulation of developmental processes, an important role in phenotypic and physiological adaptability, and the ability of plants to survive and reproduce in a changing environment. Plant DNA methylation takes place in the sequences CG, CHG, and asymmetric CHH (H symbol for A, C, or T), each with different "genetic requirements" for conservation. Asymmetric methylation of CHH cannot be maintained and requires de novo methylation in every cell cycle. The process by which this is accomplished is DNA-directed methylation with RNA (RdDM). Two plant-specific RNA polymerases, Pol IV and Pol V, synthesize small interfering RNA and long non-coding RNA molecules, important for RNA-DNA recognition. The DDR complex is a protein complex that is thought to be responsible for bringing the Pol V enzyme to the site of gene regulation, but the mechanism of specificity is unclear. Our preliminary results indicate that the MATH-BTB proteins from the manual interact with the DDR complex, and thus may mediate the positioning of the RdDM system. Recent findings indicate that BPM protein substrates are transcription factors. Proteins with such interaction capabilities have the potential to maintain a balance between methylation (mediated by Pol V) and transcription (mediated by Pol II). The main goal of the project is to elucidate the functional role of BPM proteins in the regulation of transcription and RdDM as well as possible roles in plant embryogenesis and adaptation to stress conditions. Particular attention will be paid to the identification of genes whose transcription or RdDM methylation is balanced by the BPM1 protein (chromatin immunoprecipitation-ChIP, ChIP-sequencing). Furthermore, the methylation pattern (bisulfite sequencing) of the selected genes will be associated with their expression (Q-PCR), and with embryogenesis and plant response to stress in lines with altered RdDM system or BPM gene expression.

Ključne riječi

Metilacija DNA, RdDM, MATH-BTB, embriogeneza, stres, biljke

DNA methylation, RdDM, MATH-BTB, embryogenesis, stress, plants

Znanstveno-istraživački projekti

PHYTOMETHDEV

IP-2016-06-6229

01.03.2017

28.02.2021

nije evidentirano

HRK 1.000.000

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