H2020Индивидуална стипендия2017–2019

QAPs · G-Quadruplex-associated proteins (QAPs) and their role in transcriptional regulation

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

Период: 2017-04-01 → 2019-03-31
Финансиране от ЕС: 183 455 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Протеините, които се свързват със специални четиринишкови структури в ДНК (G-квадруплекси), се изучават чрез химически сонди и секвениране. Това помага за разбирането на начина, по който се регулират гените и как се развиват раковите и неврологичните заболявания.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

G-Quadruplex-associated proteins (QAPs) and their role in transcriptional regulation

Structure dictates function in many aspects of biology. In addition to the well-known DNA double-helix made from A, C, T and G nucleotides, sequences rich in G have the potential to fold into four-stranded, alternative structures known as G-quadruplexes (G4s). G4s were initially considered a structural curiosity, but recent evidence suggests their involvement in key genome functions with strong links to neurological and cancer disease. The human genome only codes for a small fraction of sequences with the capability to fold into G4s and of these even less actually form the structure. These G4s have been located to places in the genome known to regulate how genes are expressed (called promoters) particular for genes that are very active. However, very little is known about what proteins might bind these G4 structures to control their formation and how they might regulate genome function through recruiting and modulation of the activity of critical cellular machinery. The objective of this project is to identify and study these so-called Quadruplex-associated proteins (QAPs) in a normal cellular environment by employing small chemical probe molecules and antibodies to isolate QAPs bound to G4s in conjunction with the latest quantitative proteomics and next-generation genome sequencing. The study of G4s and their native binding partners is an urgent priority to gain fundamental insights into the cellular regulation and role of G4s in genome function and to advance the understanding of G4-related disease biology.

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

Цел на проекта

In the human genome, many G-rich sequences have the potential to form 4-stranded non-canonical secondary structures known as G-quadruplexes (G4s). G4 structures have been visualized in nuclei by immunofluorescence and mapped genome-wide to regulatory chromatin regions linked to elevated transcription using G4 chromatin immunoprecipitation and high-throughput sequencing (G4 ChIP-seq). G4s are implicated in gene regulation, DNA replication and genome instability, yet little is known about their protein interaction partners or role in regulating genome activity. We therefore propose to develop G4 Rapid Immunoprecipitation Mass Spectrometry of Endogenous Proteins (G4 RIME) methodologies to elucidate the G4 interactome within a native chromatin context. ChIP-seq of protein candidates coupled with G4 ChIP-seq will verify binding at endogenous G4 structures. Using established biophysical and biochemical techniques, we will investigate binding modes, recruitment and effects on G4 dynamics. Consecutive chromatin immunoprecipitation steps using antibodies targeting characteristic chromatin marks as well as G4 ChIP-seq supported by bioinformatics analysis of genomic databases will identify proteins involved in transcriptional regulation. We will then test the consequence of stabilising G4s with small molecule ligands on the G4 interactome and how this is modulated during genomic stress. Cells genetically deficient in G4-interacting proteins will enable detailed exploration of G4 formation and transcriptional effects. This highly interdisciplinary project will provide a wide range of excellent training opportunities exploiting the Balasubramanian group’s unique position with laboratories at the Cancer Research UK Cambridge Institute (CRUK CI) and the Department of Chemistry. Overall, the studies will generate novel functional insights in the G4 biology that may ultimately lead to the identification of mechanisms and pathways for new anti-cancer agents.

Оригинален текст от CORDIS (на английски).

Участници

THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE · CAMBRIDGEКоординаторОбединеното кралство