SMT OF TFS

Quantifying transcription factor search mechanism by three-dimensional single molecule tracking

 Coordinatore UNIVERSITA VITA-SALUTE SAN RAFFAELE 

 Organization address address: Via Olgettina 58
city: MILANO
postcode: 20132

contact info
Titolo: Dr.
Nome: Maria Rosa
Cognome: Pedrazzi
Email: send email
Telefono: +39 02 2643 4845
Fax: +39 02 26434717

 Nazionalità Coordinatore Italy [IT]
 Totale costo 181˙084 €
 EC contributo 181˙084 €
 Programma FP7-PEOPLE
Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013)
 Code Call FP7-PEOPLE-2010-IIF
 Funding Scheme MC-IIF
 Anno di inizio 2012
 Periodo (anno-mese-giorno) 2012-04-16   -   2014-04-15

 Partecipanti

# participant  country  role  EC contrib. [€] 
1    UNIVERSITA VITA-SALUTE SAN RAFFAELE

 Organization address address: Via Olgettina 58
city: MILANO
postcode: 20132

contact info
Titolo: Dr.
Nome: Maria Rosa
Cognome: Pedrazzi
Email: send email
Telefono: +39 02 2643 4845
Fax: +39 02 26434717

IT (MILANO) coordinator 181˙084.00
2    FONDAZIONE CENTRO SAN RAFFAELE DEL MONTE TABOR

 Organization address address: Via Olgettina 60
city: MILANO
postcode: 20132

contact info
Titolo: Dr.
Nome: Maria Rosa
Cognome: Pedrazzi
Email: send email
Telefono: +39 02 26432729
Fax: +39 02 26432752

IT (MILANO) participant 0.00

Mappa


 Word cloud

Esplora la "nuvola delle parole (Word Cloud) per avere un'idea di massima del progetto.

tfs    living    single    cells    stress    tracking    mechanisms    tf    cell    plan       search    sliding    efficient    limitations    transcription    quantify    molecule    dna    fractal    smt    microscopy    diffusion    compact    exploration    individual   

 Obiettivo del progetto (Objective)

'Transcription factors (TFs) are capable to identify their target sites on DNA in a extremely efficient fashion. The observation that a simple 3D diffusion search cannot account for such efficient targeting has led to different mechanisms to be proposed in the last decades, including compact exploration induced by facilitated diffusion (i.e. combination of 1D sliding and 3D diffusion) or by fractal-like chromatin structure. Despite the evidence for 1D sliding of TFs on DNA and for fractal nuclear organization, it still needs to be demonstrated that these phenomena play a significant role in the TF search mechanism in a living cell. Fluorescence microscopy and in particular single molecule tracking (SMT) provides an excellent tool to investigate the kinetics of proteins in living samples. SMT has mostly been applied to in-vitro and in-membrane environments due to limitations associated to prolonged 3D tracking. In this project we plan to overcome the limitations associated to 3D SMT and to apply the developed technology to investigate the in-vivo search mechanisms of p53, an important TF, involved in the determination of the cell fate under stress conditions. We aim to quantify the role of compact exploration in p53 targeting in living cells, and by the analysis of mutated p53 how 1D sliding affects such phenomenon. Furthermore, we plan to identify the changes in the p53-DNA association and in the p53 search after induction of cell stress by DNA damage. Summarizing, the project aims to strongly improve current methods for tracking individual molecules in 3D and to advance our knowledge about the mechanisms of TF targeting.'

Introduzione (Teaser)

Single-molecule approaches, recently empowered by advanced microscopy techniques, are predicted to become more widespread in subsequent years. Using single-molecule microscopy, it was possible to reliably quantify the movement and interactions of individual transcription factors in the nucleus of living cells.

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