H2020Индивидуална стипендия2015–2017

InterAcTEV · Genome-wide analysis of RNA and protein interacting profiles during a plant virus infection

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2015-05-01 → 2017-04-30
Финансиране от ЕС: 170 122 €
Участници: 1
Схема: MSCA-IF-EF-RI

Линиите свързват координатора с партньорите.

Накратко на български

Взаимодействията между вирусни протеини и РНК в растенията се анализират, за да се открият слаби места в защитата на вирусите. Това помага за разработването на методи за защита на посевите и осигуряване на хранителните ресурси.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Genome-wide analysis of RNA and protein interacting profiles during a plant virus infection

Viruses constitute a major class among plant pathogens. RNA viruses encompass the majority of plant viruses, accounting for substantial losses in crop yields worldwide. Understanding how viruses incite diseases and how plants defend themselves against them is critical to develop effective antiviral approaches for crop protection that ensure food availability in the following years. Plants have evolved two highly specific adaptive immunity pathways to face viruses, viroids and other pathogens. First, RNA silencing is an ancient antiviral mechanism induced by viral double-stranded RNAs (dsRNAs) that are processed into 21 to 24 nt small RNAs (sRNAs). In the antiviral RNA-silencing pathway model (see Figure), virus-derived sRNAs (vsRNAs) associate with a plant ARGONAUTE (AGO) protein, and sRNAs guide the AGO to interact and repress viral RNAs sharing sequence complementary with them. Second, plants have evolved involves disease resistance (R) proteins that recognize viral-encoded proteins through direct protein-protein interactions or through an indirect effect due to the interaction of a viral protein with a host protein included in a larger complex containing an R protein. This project intended to better understand the molecular interplay occurring during plant/virus interactions. Our main goal was to identify new viral targets to develop effective antiviral resistance for crop protection. In particular, we wanted to identify AGO target sites highly susceptible to artificial sRNA-mediated inactivation, as well as plant proteins interacting with viral proteins upon infection. To accomplish these objectives, first we explored the possibility of using amiRNAs and syn-tasiRNAs to specifically interfere with infections by economically important viruses and viroids. The combined use of recent high-throughput methods for artificial sRNA construct generation and of the Nicotiana benthamiana/Tomato spotted wilt virus (TSWV) and the N. benthamiana/Potato spindle tuber viroid (PSTVd) pathosystems allowed for the simple and time-effective screening of multiple artificial sRNAs targeting sites distributed along TSWV and PSTVd RNAs. We have identified several AGO target sites in TSWV and PSTVd RNAs that are efficiently targeted by specific amiRNAs or syn-tasiRNAs. Second, we have further optimized our RNA immunoprecipitation followed by high-throughput sequencing (RIP-Seq) approach. Our current methodology has already identified potential new AGO1 target transcripts that are currently being investigated, and will serve to detect AGO target sites in viral RNAs which could be used as ideal targets for artificial sRNA-mediated inactivation. And third, we have used the well-characterized N. benthamiana/Tobacco etch virus (TEV) pathosystem to analyse the protein interactors of several TEV proteins. Several TEV infectious clones including TEV proteins tagged with the Twin-Strep tag (TST) system have been generated and are being analyzed.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Plant (+)strand RNA viruses account for important losses in crop yields worldwide. They complete complex infectious cycles by replicating, moving and inactivating plant defense mechanisms through multiple interactions of their RNA or proteins with host factors. Plants have evolved two main antiviral immunity systems operated by ARGONAUTE (AGO) proteins that target viral RNAs in the RNA silencing pathway or by resistance proteins that recognize viral-encoded proteins through protein-protein interactions. However, neither the identification of AGO target sites in viral RNAs nor the elucidation of the entire repertoire of proteins interacting with viral proteins has been reported for any plant virus. Now, the recent development of new technologies (and associated tools) such as high-throughput DNA sequencing and high performance mass spectrometry (MS) platforms makes possible to address these questions at a genome-wide level. InterAcTEV will combine these new technologies to study the recently described Arabidopsis/Tobacco etch virus (TEV)-TAMPS pathosystem with a multidisciplinary approach aimed to identify AGO target sites in TEV RNAs and to generate the first complete protein-protein interactome map for a plant virus. First, RNA-immunoprecipitation coupled with high-throughput sequencing methods will be optimized to identify AGO target sites in TEV RNAs at a genome-wide level. Second, we will combine novel high-affinity purification methods with MS analyses to identify the specific pool of Arabidopsis proteins interacting with each TEV protein. InterAcTEV will generate key knowledge to open new avenues for the understanding of resistance mechanisms in plants and to develop novel antiviral strategies for crop protection and securement of world’s food supply. Finally, the funding of InterAcTEV will allow the Experienced Researcher to initiate his own research project in the host laboratory and facilitate his transition to an independent research position in Europe.

Оригинален текст от CORDIS (на английски).

Участници

AGENCIA ESTATAL CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS · MadridКоординаторИспания