H2020Индивидуална стипендия2017–2019

MuRChap · The mutation-buffering capacity of RNA chaperones

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

Период: 2017-09-01 → 2019-08-31
Финансиране от ЕС: 183 455 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

РНК шапероните, като протеина CYT-19, помагат на неправилно сгънатите РНК молекули да възвърнат правилната си форма. Това помага да се разбере как клетките компенсират вредните мутации и колко стабилно е сгъването на РНК в живите организми.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

The mutation-buffering capacity of RNA chaperones

Due to their intrinsic thermodynamic properties, RNA can misfold easily in cells. One way to mitigate RNA misfolding is through the actions of RNA chaperones, which bind and unwind structured RNA molecules and thereby offer opportunities for these misfolded species to refold properly. Such rescue activity has implications for the fitness effects of individual mutations - at least mutations that compromise RNA folding or structure might be buffered by RNA chaperones. However, little is known about the rules governing such mutation buffering. Here, we describe how a model RNA chaperone, the DEAD-box RNA helicase CYT-19, affects the fitness effects of mutations in a model structured RNA, the Tetrahymena group I intron, whose self-splicing activity is dependent on its structure. The goal was to comprehensively catalogue mutational effects on this self-splicing intron in the absence and presence of CYT-19. Conclusions: To date, this work has yielded considerable information about the mutational effects on the Tetrahymena group I intron. My results highlighted the overall complexities in delineating such mutational effects on a model RNA. More importantly, understanding the in vivo mutational effects on a model RNA will have considerable implications for understanding the robustness in RNA folding. Through this project, two-way transfer of knowledge and skills has been achieved.

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

Цел на проекта

The goal of this project is to investigate how mutations that affect RNA structure can be buffered in trans by RNA chaperones using a combination of experimental and computational approaches. The Warnecke lab (the host) recently showed that RNA chaperones, like their protein chaperone counterparts, can buffer the fitness effects of deleterious mutations in Escherichia coli (Rudan et al. 2015 eLife, 4:e04745). However, the rules governing mutation buffering at the RNA level remain poorly understood. Do RNA chaperones rescue misfolded RNA intermediates? Do they alleviate the effects of mutation that lead to excessively stable secondary structures? Which mutations are amenable to buffering and which are not? And does that presence of RNA chaperones render their substrate RNAs more evolvable? Here, I will evaluate the buffering capacity of a model RNA chaperone, the DEAD-box RNA helicase CYT-19, by exploring how it affects the mutational robustness of the Tetrahymena group I intron, whose self-splicing activity is dependent on its structure. Following systematic site-directed and random mutagenesis, I will assay differential splicing activity of the generated intron variants and compare results to predictions from RNA structural modelling. Importantly, I will assay activity both in the presence and in the absence of CYT-19 to identify mutations that are buffered by RNA chaperone activity. To further understand the structural impact of chaperone-dependent mutations, I will use in-cell SHAPE-Seq to determine the mutated intron structures. To my knowledge, this is the first quantitative assessment of mutational effects on RNA in the presence of an RNA chaperone. The expected outcome will improve our understanding of RNA robustness, and may reveal insights into making better RNA-based tools.

Оригинален текст от CORDIS (на английски).

Участници

IMPERIAL COLLEGE OF SCIENCE TECHNOLOGY AND MEDICINE · LondonКоординаторОбединеното кралство