H2020Индивидуална стипендия2021–2023

MolecularArp · Molecular analysis of the properties of human Arp2/3 complex family members

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

Период: 2021-04-01 → 2023-03-31
Финансиране от ЕС: 212 934 €
Участници: 1
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Семейството от Arp2/3 комплекси в човека се анализира, за да се разбере как различните му видове влияят върху стабилността на актиновите нишки. Това помага за разбирането на процеси като клетъчното движение и развитието на човешките 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

Molecular analysis of the properties of human Arp2/3 complex family members

Arp2/3 has been known for over two decades, yet many questions remain regarding its regulation and the disassembly of actin branches generated by the complex. Recent breakthroughs have revealed that Arp2/3 in humans is actually a family of 8 complexes with distinct properties, which has important implications for human development and disease. However, a full understanding of why these complexes differ and whether they have unique cellular functions is still lacking. To address these outstanding questions, the project utilized a combination of in vitro biochemical approaches with defined recombinant Arp2/3 complexes and cell-based assays using CRISPR-Cas9 genome edited cells in combination with advanced live cell imaging. The project has achieved most of its objectives. During the project, it has been discovered that different type I Nucleation Promoting Factors (NPFs) can regulate Arp2/3 iso-complexes similarly, but type II NPFs regulate them differently, indicating that specific Arp2/3 iso-complexes are involved in different cell signalling pathways. Additionally, the fellow’s study on the dissociation of Arp2/3-mediated branches revealed that actin nucleated by various Arp2/3 iso-complexes has different stability, which contributes to understanding the regulation of actin dynamics through different Arp2/3 iso-complexes. However, due to difficulties in obtaining purified MICAL2, it was not examined how it promotes the rapid debranching of Arp3B-containing complexes. Although the analysis of the cellular function of individual Arp2/3 family members is still ongoing, the preliminary results suggest that ArpC5-containing Arp2/3 complexes play an important role in cell migration. Further work is needed to fully understand the cellular functions of Arp2/3 iso-complexes. Overall, this project has achieved most of its objectives. The scientific outputs have contributed significantly to our molecular understanding of Arp2/3 iso-complexes mediated actin nucleation and dissociation. During the project, the researcher was able to share their knowledge of biochemistry and molecular biology with the host lab and gained valuable expertise in cell biology. Moreover, the communication and collaboration established during the project have benefited the wider scientific community.

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

Цел на проекта

The actin cytoskeleton is a dynamic polymer system, that provides the driving force and structural support for the physical integrity of cells, and their interactions. It also regulates many essential cellular processes, such as membrane trafficking and cell migration. Not surprisingly, its malfunction results in a wide variety of syndromes and diseases. Branched actin networks can only be assembled by the Arp2/3 complex, consisting of seven protein subunits. Activated by nucleation promoting factors (NPFs), Arp2/3 binds to the side of an actin filament to initiate an actin branch and new filament growth. Arp2/3 has been regarded as a single entity, but in mammals, three of its subunits exist as two isoforms, thus allowing the formation of 8 different complexes. Recent work demonstrates that the 8 human Arp2/3 complexes have unique cellular properties. However, it is unclear why these complexes are so different and whether they have distinct cellular functions. Using in vitro biochemical assays and single molecule analysis together with sophisticated cell biology approaches, I will determine the molecular basis for the individual properties of the different Arp2/3 complexes. Observing in vitro reconstituted actin networks, I will perform a detailed analysis of the actin nucleation properties of the 8 Arp2/3 complexes and the impact of different NPFs. In addition, I will also analyse actin branch disassembly to uncover the molecular basis of actin debranching, which still remains to be established. To investigate the cellular function of individual Arp2/3 family members, I will generate genome edited cells expressing defined Arp2/3 complexes and examine their lamellipodial actin dynamics and cell migration. Moreover, by generating Arp2/3 complex specific biosensors, I will map the cellular distribution of individual Arp2/3 complexes. My combined approaches will allow me to understand the molecular and cellular basis of Arp2/3 diversity in humans.

Оригинален текст от CORDIS (на английски).

Участници

THE FRANCIS CRICK INSTITUTE LIMITED · LondonКоординаторОбединеното кралство