Descrizione del progetto
Meccanismi fondamentali della replicazione dei cromosomi
La duplicazione fedele e la trasmissione di informazioni genetiche ed epigenetiche all’interno delle cellule viventi sono condotte da grandi complessi macromolecolari noti come replisomi, che coordinano gli eventi enzimatici durante la duplicazione dei cromosomi. Il progetto REPLISOMEBYPASS, finanziato dall’UE, intende sviluppare un approccio di imaging a singola molecola per studiare i meccanismi con cui i replisomi mantengono l’efficienza e la coordinazione durante le collisioni con gli ostacoli sul cromosoma. Il progetto si avvale di un approccio di imaging multidimensionale a singola molecola di recente realizzazione per visualizzare in tempo reale la coordinazione durante la replicazione. L’obiettivo è comprendere le conseguenze delle collisioni con il replisoma e i meccanismi che consentono di aggirare o innescare il collasso della forca di replicazione. Il lavoro svolto nell’ambito del progetto permetterà di scoprire l’origine molecolare del danno cromosomico alla base di numerose patologie.
Obiettivo
Faithful duplication and transmission of genetic and epigenetic information is the most vital cellular
function for the preservation and proliferation of life. In cells, this process is conducted by large
macromolecular complexes, known as replisomes, that coordinate the sequence of enzymatic events
during chromosome duplication. While recently developed single-molecule techniques promise
unprecedented access to the complex inner workings of these sophisticated machines, most studies
conducted have focused on individual factors, operating on non-physiological substrates, which has
provided an incomplete molecular picture.
My recent development of a multidimensional, single-molecule imaging approach that allows
for real-time visualisation of coordination during replication represents a significant breakthrough
in our ability to study macromolecular machines in vitro. Building on this success, here I describe
single-molecule imaging approaches to address one of the long-standing questions in chromosome
biology: How do replisomes maintain efficiency and coordination during collisions with obstacles
on the chromosome?
Our objective is to develop a complete molecular understanding of the consequences of
replisome collisions and the underlying mechanisms that allow for bypass or trigger replication fork
collapse. We will begin this long-term research effort by addressing several issues fundamental to
chromosome replication: How does replisome coordination and composition change during
encounters with topological barriers in chromosomes? What are the dynamic events that underlie
nucleosome disassembly by histone chaperones during replication? How does the eukaryotic
replisome collaborate with histone chaperones to ensure faithful inheritance of epigenetic
information encoded on histones?
These studies will provide a framework for understanding the dynamics of replisome collisions
and the molecular origin of chromosome damage underlying many diseases.
Campo scientifico
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-STG - Starting GrantIstituzione ospitante
80539 Munchen
Germania