H2020Индивидуална стипендия2021–2023

NTON · Unravelling the mechanism of nuclear transport using optical nanopores

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

Период: 2021-08-01 → 2023-07-31
Финансиране от ЕС: 175 572 €
Участници: 1
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Ядрените пори контролират движението на молекули, като например пропускат малки частици, но изискват специални протеини за пренасяне на големи молекули. Разбирането на този механизъм помага да се обясни появата на рак и невродегенеративни 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

Unravelling the mechanism of nuclear transport using optical nanopores

The nuclear pore complex (NPC) is the gatekeeper of the nucleus that regulates the flow of all molecules across the nuclear envelope. Remarkably, this ~40 nm wide pore is capable of efficient and fast transport while remaining highly selective. Its dense central channel is composed of a highly dynamic spaghetti-like mesh of intrinsically disordered proteins that allows small molecules to pass freely, whereas large macromolecules (>40 kDa) rely on specific transporter proteins that ferry their cargo across the pore. This process is vital to the well-being of the cell, as the NPC poses a gate between the nucleus, the place of the genetic material and mRNA transcription, and the cytosol, where proteins are synthesized from mRNA by ribosomes - hence posing a bottleneck for the central dogma of molecular biology by regulating the flow of mRNA. At the same time, the NPC protects the genetic material from viral intruders, but needs to allow efficient import of proteins to the nucleus to maintain the structural integrity of the genome, and perform and regulate gene transcription. Given its vital importance, it is unsurprising that mutations of the NPC have been linked to numerous neurodegenerative diseases and various forms of cancer. Even though various models have been proposed, the fundamental biophysical mechanism of nuclear transport and of the selectivity of the NPC has not been resolved yet. One of the main reasons that have hindered our progress in understanding nuclear transport is the lack of experimental techniques that can probe the structure and dynamics of the disordered proteins and transport receptors inside the NPC channel and during transport with sufficient spatiotemporal resolution. In this project, we have combined nanotechnology with single-molecule fluorescence microscopy to build and study biomimetic, minimal versions of the NPC.

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

Цел на проекта

The nuclear pore complex (NPC) is the gatekeeper of the nucleus that regulates the flow of all molecules across the nuclear envelope. Remarkably, this ~40 nm wide pore is capable of efficient and fast transport while remaining highly selective. Its dense central channel is composed of a highly dynamic spaghetti-like mesh of intrinsically disordered proteins that allows small molecules to pass freely, whereas large macromolecules (>40 kDa) rely on specific transporter proteins that ferry their cargo across the pore. Even though various models have been proposed, the fundamental biophysical mechanism of nuclear transport and of the selectivity of the NPC has not been resolved yet. One of the main reasons that have hindered our progress in understanding nuclear transport is the lack of experimental techniques that can probe the structure and dynamics of the disordered proteins and transport receptors inside the NPC channel and during transport with sufficient spatiotemporal resolution. In the proposed action, I aim to establish a modifiable biomimetic platform for single-molecule investigations of nuclear transport. This interdisciplinary and innovative approach combines solid-state nanopores, coated with disordered proteins to mimic the nuclear pore, with optical detection in zero-mode waveguides and DNA origami nanotechnology. The proposed experiments will allow to monitor the translocation of single fluorescently labelled molecules through biomimetic NPCs with excellent sensitivity and specificity. I will use this platform to elucidate the biophysical principles that underlie the structure and selectivity of the NPC channel and the mechanism of active nuclear transport. The direct insights on the single-molecule level obtained by the results of this project will be crucial for our understanding of the molecular principles that govern nuclear transport.

Оригинален текст от CORDIS (на английски).

Участници

TECHNISCHE UNIVERSITEIT DELFT · DelftКоординаторНидерландия