H2020Индивидуална стипендия2017–2019

TF3C_EM · Structure-function studies of the general transcription factor IIIC (TFIIIC)

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

Период: 2017-03-01 → 2019-02-28
Финансиране от ЕС: 159 461 €
Участници: 1
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Структурата и работата на протеиновия комплекс TFIIIC се анализират, за да се разбере как се започва четенето на гените за транспортна РНК. Познаването на този механизъм помага за разбирането на заболявания като рака на яйчниците.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Structure-function studies of the general transcription factor IIIC (TFIIIC)

During protein synthesis, tRNAs are required for delivery of the correct amino acids to the growing peptide chain during translation of the messenger RNA. Misregulation in the transcription of tRNA genes is often associated with cancer in humans, especially in ovarian tumors. Studying the basic transcription mechanism thus has a direct health implication for society in the long-term. ‘TF3C_EM’ focused on characterizing the process of transfer RNAs (tRNAs) gene transcription structurally. In eukaryotes, the task of transcription is shared by three types of RNA polymerases, selected depending upon the gene being read. tRNA gene (tDNA) transcription begins with the recruitment of the multi-protein transcription factor IIIC (TFIIIC) to two characteristic gene-internal promoter elements named A- and B-boxes. This sets the stage for the assembly of the pre-initiation complex which also includes transcription factor IIIB and RNA polymerase III and thereby, the start of transcription. However, the lack of information on the overall TFIIIC structure and its DNA binding mechanism has hampered the understanding of transcription initiation. ‘TF3C_EM’ focused on the structure-function elucidation of TFIIIC with the following objectives: (1) Characterization of the molecular architecture of TFIIIC, (2) Understanding the tDNA recognition mechanism by TFIIIC and (3) Understanding the dynamics between the two subdomains of TFIIIC- τA and τB. I obtained a low resolution structure of TFIIIC, purified from yeast using the TAP-tag strategy. I was also able to study the complete pre-initiation complex, paving the way for further understanding the transcription initiation. Moreover, ‘TF3C_EM’ gave me an opportunity to widen my skill set from working with different model organisms (yeast, insect cells), to handling high-end cryo-electron microscopes, data collection and processing. I was also able to build on my project management and science communication skills at EMBL during this time. These skills are extremely valuable for my future position.

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

Цел на проекта

Transfer RNAs (tRNAs) act as adaptor molecules, decoding messenger RNA and delivering the correct amino acid to the growing peptide chain. Being instrumental in the cellular translation machinery, it is important to understand their synthesis mechanism. In eukaryotes, tRNA gene (tDNA) transcription begins with transcription factor IIIC (TFIIIC) bound to 2 gene-internal promoter elements, A- and B-box. This is followed by the recruitment of transcription factor IIIB and RNA polymerase III to form the transcription pre-initiation complex. In ‘TF3C_EM’, I will focus on the mechanism and molecular architecture of the very first step, i.e., TFIIIC binding to tDNA. TFIIIC is composed of two subcomplexes, τA and τB, that bind respectively to the A- and B-box, and are flexibly linked to accommodate varying lengths between their binding sites. Interestingly, TFIIIC has recently been termed as ‘guardian of the genome’ in light of its extra transcriptional roles, such as chromatin remodelling. This further highlights the importance of exploring its underlying structure. Using recombinantly purified TFIIIC, I will characterise the fully assembled TFIIIC complex and TFIIIC-tDNA interactions in vitro. To this end, an inter-disciplinary approach will be used, spanning complementing biophysical and structural techniques of single molecule FRET, cryo electron microscopy and X-ray crystallography. Thus, in my project ‘TF3C_EM’, I will tackle 3 major objectives: (1) characterisation of the molecular architecture of TFIIIC, (2) understanding the tDNA recognition mechanism by TFIIIC and (3) understanding the dynamics between τA and τB. It will unravel the structure-function correlation of TFIIIC and pave the way for further understanding the transcription initiation by Pol III. At the same time, it will allow me to pursue research in an unparalleled scientific environment and significantly contribute to shape my future career as an independent researcher.

Оригинален текст от CORDIS (на английски).

Участници

EUROPEAN MOLECULAR BIOLOGY LABORATORY · HeidelbergКоординаторГермания