PLANMIP · Investigation of the plant miRNA pathway
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2011-09-01 → 2015-08-31
- Финансиране от ЕС
- 100 000 €
- Участници
- 1
- Схема
- MC-CIG
Линиите свързват координатора с партньорите.
Накратко на български
Растителните микроРНК и протеинът AGO1 се изследват, за да се разбере къде точно в клетъчната мембрана се намират и как функционират. Това помага да се разбере как се регулират гените в растенията и какви процеси влияят върху тях.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Periodic Report Summary 1 - PLANMIP (Investigation of the plant miRNA pathway)
The project objectives were to provide answers to the following questions: 1. What is the membrane compartment(s) to which AGO1 and miRNAs localize? 2. What is the function of membrane-anchored RISC, and how is RISC recruited to membranes? 3. What specific genes influence inhibition of the miRNA pathway following inhibition of isoprenoid biosynthesis? In the four months that have elapsed from project start to project termination (due to the award of an ERC Starting Grant to the researcher) the following work and results have been obtained: AGO1 localization We have shown that the N-terminal domain alone is sufficient to drive GFP reporter localization to distinct cytoplasmic bodies whose identity remains unknown to us. We have used westerns to identify tissues where AGO1 is particularly highly expressed, and are continuing our efforts on immunofluorescence and elctron microscopy on those tissues. AGO1 membrane recruitment a) Construction of HPB-AGO1 The construct had still not been completed by 31/12/2012 despite employment of different strategies. Work is continuing to get this important construct made. ago1-3/mcca mutant lines have been selected and are ready for transformation. b) AGO1 yeast two-hybrid screening. The constructs have been made and shown to be expressed as Gal4 fusions in yeast by western analysis. Plating on selective media has shown that they do not cause reporter autoactivation. A screen has been succesfully conducted with the N-terminal domain of AGO1, and a single colony positive for activation of all three reporters has been recovered. Work is ongoing to characterize this by retransformation, sequencing etc. c) ago1 suppressor screens We have introduced a miR398 sensor into ago1-38 by crossing, but have not yet carried out the mutagenesis. Isoprenoid/miRNA crosstalk This part of the project was only initiated by ordering and growing Arabidopsis ecotypes during the four months covered here. The potential impact of the expected results of the project reaches far. The links between membrane and isoprenoid biology on the one side and RNA silencing on the other remain nearly fully unexplored, yet our recently published results clearly demonstrate the existence of such links (Brodersen et al. (2012), PNAS 109, 1778-83). For example, the recent demonstration that inhibition of miR-33 in primates influences serum levels of HDL and VLDL triglycerides in conjunction with our demonstration of miRNA pathway inhibition by statins could shed some light on atheroprotective action on this very widely used group of drugs.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
MicroRNAs (miRNAs) are 20-22 nt non-coding RNAs that regulate gene expression post transcriptionally via base pairing to complementary target mRNAs. They have fundamental importance for development and stress adaptation in plants and animals. Although a molecular frame work for miRNA biogenesis, degradation and action has been established, many aspects of this important gene regulatory pathway remain unknown. This project explores four main points. First, we propose to use genetic approaches to identify factors required for translational repression by miRNAs in plants. This mode of action was until recently thought to occur only exceptionally in plants. My post doctoral work showed that it occurs in many miRNA-target interactions. The mechanism remains unknown, however, leaving open a fertile area of investigation. Second, we wish to test specific hypotheses regarding the in vivo role of miRNA mediated endonucleolysis of mRNA targets. Long believed to serve exclusively as a degradation mechanism, we propose to test whether this process could have important functions in biogenesis of long non-coding RNA derived from mRNAs. Third, my postdoctoral work has provided unique material to use molecular genetics to explore pathways responsible for miRNA degradation, an aspect of miRNA biology that only now is emerging as being of major importance. Finally, our unpublished results show that plant miRNAs and their associated effector protein Argonaute (AGO) are associated with membranes and that membrane association is crucial for function. This is in line with similar data recently obtained from different animal systems. We propose to use genetic, biochemical and cell biological approaches to clarify to which membrane compartment AGO and miRNAs are associated, how they are recruited to this compartment, and what the precise function of membrane association is. These innovative approaches promise to give fundamental new insights into the inner workings of the pathway.
Оригинален текст от CORDIS (на английски).
Участници
- KOBENHAVNS UNIVERSITET · KOBENHAVNКоординаторДания
Връзки
Данни: CORDIS, © Европейски съюз