H2020Индивидуална стипендия2022–2024

Iso-Proline CTD · Hidden in plain sight: Masking RNA Pol II phosphorylation via proline isomerization during gene expression

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

Период: 2022-11-15 → 2024-11-14
Финансиране от ЕС: 219 312 €
Участници: 1
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Регулацията на генната експресия се изследва чрез структурни промени в протеиновия комплекс, който синтезира шаблоните за всички протеини в тялото. Разбирането на тези механизми помага да се разбере как се контролира производството на протеини и какво се случва при заболявания като рака.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Hidden in plain sight: Masking RNA Pol II phosphorylation via proline isomerization during gene expression

Over 15,000 different proteins are produced across all tissue types in the human body, each with its own timing of expression, specific cell-type localization, and size. Remarkably, the templates for all these proteins are synthesized by a single protein complex. Given that many types of cancer arise from dysfunctions in protein production, understanding how the synthesis of these products is precisely orchestrated becomes essential to our understanding of human disease. The focus of this project is to capitalize on recent technological advances to introduce a new layer of gene expression regulation, previously hindered by the lack of essential tools. For decades, our field has wondered how the remarkable specificity of gene expression is achieved. Research has concluded that certain chemical modifications displayed on the above-mentioned protein complex function as “active/inactive” signals. The cell reads permutations of these signals together, creating a “code” of gene expression. Our results introduce a new variable to this code—structural changes that occur without the addition of chemical modifications, essentially “hidden in plain sight.” Taking advantage on these insights, we have generated an extensive dataset through novel technological approaches, which collectively is helping us understand how gene expression is regulated and what occurs in cells when expression goes awry in disease.

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

Цел на проекта

Prolines are unique amongst aminoacids given that they naturally exist in two isomeric states: CIS and TRANS. The transition between these states is slow but can be catalyzed by the activity of proline isomerases such as PIN1. Recent in vitro work suggests that PIN1 can alter the phosphorylation dynamics of the C-terminal domain (CTD) of RNA polymerase II (Pol II) by inhibiting phosphatase recognition. Given Pol II is solely responsible for transcribing all protein-coding genes and that CTD phosphorylation dictates the timing of RNA co-transcriptional processes, these observations suggest a crucial role for CTD proline isomerization in gene expression. Importantly, mutations in PIN1 are associated with cancer progression but a direct role for proline isomerization has remained understudied given the technical limitations imposed by its complex enzymology, such as the inability of differentiating CIS and TRANS isomers using Mass Spectrometry. In this project, I aim to functionally dissect the role of proline isomerization during Pol II transcription using rigorous biochemical, cellular and genomic techniques. Specifically, I will exploit new technological advances in peptide synthesis to permanently “lock” prolines in CIS or TRANS and identify novel isomer-specific interacting factors. I will systematically examine the consequences of CTD proline mutations and of rapid depletion of PIN1 in human cells, focusing on CTD-dependent co-transcriptional RNA processes such as splicing and poly-A–dependent 3’ termination. At the basic research level, my results will provide unprecedented resolution to the dynamics of Pol II phosphorylation, which underlies regulation of gene expression in multicellular organisms. Translationally, given that various cancers hijack the transcriptional programmes of the cell, this mechanistic understanding of CTD proline isomerization will better equip future clinical studies interested in the yet-to-be-characterized role of PIN1 in oncogenesis.

Оригинален текст от CORDIS (на английски).

Участници

KOBENHAVNS UNIVERSITET · KOBENHAVNКоординаторДания