Forum für Wissenschaft, Industrie und Wirtschaft

Hauptsponsoren:     3M 
Datenbankrecherche:

 

Need for Speed: High-speed Measurements of Molecular Motion in the Cell Nucleus

03.12.2012
Heidelberg scientists study interactions between proteins and DNA in living cells

Using a new measurement technique, Heidelberg researchers have succeeded in tracking interactions between proteins and DNA in the cell nucleus at a resolution of 1/1000 of a second.


Microscopy images after bleaching the fluorescence in a circular or rectangular region. The dark “shadows” represent the bleached proteins that moved as the image was being recorded.

Picture credits: Fabian Erdel and Karsten Rippe

They were able to measure the binding of highly specialised protein complexes that specifically change the spatial structure of the genetic information, thereby controlling the readout of the DNA information. The work of Dr. Karsten Rippe and his team was carried out at the BioQuant Center of Heidelberg University and the German Cancer Research Center.

Their research has demonstrated that the positioning of nucleosomes – complexes of DNA and specialised proteins – is a precisely regulated molecular process. Aberrant regulation can be linked to several types of cancer. The results of these studies were published in the journal PNAS.

In the human genome, the DNA strands are wrapped around specific packaging proteins, the histones. Located between these complexes, called nucleosomes, are histone-free DNA sequences that connect the nucleosomes, much like a pearl necklace. “Activating a gene requires freely accessible DNA. If the corresponding DNA is occluded in the nucleosome, the gene is often turned off”, explains Dr. Rippe.

“Hence, the nucleosome positions determine the readout pattern of the DNA sequence. The free DNA between two nucleosomes is more easily accessible than the DNA sequences in a nucleosome.” Molecular machines called chromatin remodelers can use energy to move nucleosomes along the DNA chain. Thus they establish the readout pattern that, along with other factors, determines the active DNA programme of the cell.

Rippe’s team of scientists are using fluorescence microscopy to investigate how the chromatin remodelers control the readout of the genetic information. With it they were able to measure that most of the approx. one million chromatin remodelers in the human cell transiently bind to nucleosomes to test whether all the approx. 30 million nucleosomes are at the right position. A new way of measurement was needed to understand how these molecular machines work. “We had to record short binding events at a resolution of 1/1000 of a second and at the same time detect the rare events with a binding time of several seconds or even minutes”, says Karsten Rippe. Doctoral student Fabian Erdel came up with an idea that led to “Pixel-wise Photobleaching Profile Evolution Analysis”, or 3PEA, which can be used to take such measurements in living cells.

In his experiments, Fabian Erdel used a laser beam to extinguish the artificial fluorescent tag attached to the chromatin remodelers. He noticed that the “bleached” proteins produced a “shadow” when they moved while the image was being recorded. The shape of this shadow depended on how much the movement of the chromatin remodelers slowed down due to binding to nucleosomes. “It was not easy calculating duration times of binding from the shadow image, but it was worth the effort. Our method has exciting new applications because we can use it to measure the binding of proteins in living cells very quickly and precisely”, remarks Fabian Erdel.

Using 3PEA measurements, the researchers demonstrated that an individual chromatin remodeler travels through the entire cell nucleus within a single second testing more than 300 nucleosomes – mostly without becoming active. Only occasionally the molecular machine would bind to a nucleosome for several seconds or even minutes, causing it to shift position on the DNA. Dr. Rippe and his team next want to decode signals that activate the chromatin remodelers at certain locations on the genome.

For more information, go to http://malone.bioquant.uni-heidelberg.de.

Original publication:
F. Erdel, K. Rippe: Quantifying transient binding of ISWI chromatin remodelers in living cells by pixel-wise photobleaching profile evolution analysis, PNAS, 20 November 2012, vol. 109, no. 47, E3221-3230 (online 5 November 2012, doi:10.1073/pnas.1209579109).

Contact:
Dr. Karsten Rippe
BioQuant, phone: +49 (0)6221 54-51376
karsten.rippe@bioquant.uni-heidelberg.de

Heidelberg University
Communications and Marketing
Press Office, phone: +49 (0)6221 54-2311
presse@rektorat.uni-heidelberg.de

Marietta Fuhrmann-Koch | idw
Further information:
http://malone.bioquant.uni-heidelberg.de
http://www.uni-heidelberg.de

More articles from Life Sciences:

nachricht Quasi-sexual gene transfer drives genetic diversity of hot spring bacteria
29.05.2015 | Carnegie Institution

nachricht Scientists use unmanned aerial vehicle to study gray whales from above
29.05.2015 | NOAA National Marine Fisheries Service

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: Galapagos-Vulkanismus: Überraschend explosiv

Internationales Vulkanologen-Team präsentiert neue Erkenntnisse zur Eruptions-Geschichte

Vor 8 bis 16 Millionen Jahren gab es im Gebiet der heutigen Galapagos-Inseln einen hochexplosiven Vulkanismus. Das zeigt erstmals die Auswertung von...

Im Focus: Lasers are the key to mastering challenges in lightweight construction

Many joining and cutting processes are possible only with lasers. New technologies make it possible to manufacture metal components with hollow structures that are significantly lighter and yet just as stable as solid components. In addition, lasers can be used to combine various lightweight construction materials and steels with each other. The Fraunhofer Institute for Laser Technology ILT in Aachen is presenting a range of such solutions at the LASER World of Photonics trade fair from June 22 to 25, 2015 in Munich, Germany, (Hall A3, Stand 121).

Lightweight construction materials are popular: aluminum is used in the bodywork of cars, for example, and aircraft fuselages already consist in large part of...

Im Focus: Wie Solarzellen helfen, Knochenbrüche zu finden

FAU-Forscher verwenden neues Material für Röntgendetektoren

Nicht um Sonnenlicht geht es ihnen, sondern um Röntgenstrahlen: Wissenschaftler der Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU) haben zusammen mit...

Im Focus: Festkörper-Photonik ermöglicht extrem kurzwellige UV-Strahlung

Mit ultrakurzen Laserpulsen haben Wissenschaftler aus dem Labor für Attosekundenphysik in dünnen dielektrischen Schichten EUV-Strahlung erzeugt und die zugrunde liegenden Mechanismen untersucht.

Das Jahr 1961, die Erfindung des Lasers lag erst kurz zurück, markierte den Beginn der nichtlinearen Optik und Photonik. Denn erstmals war es Wissenschaftlern...

Im Focus: Solid-state photonics goes extreme ultraviolet

Using ultrashort laser pulses, scientists in Max Planck Institute of Quantum Optics have demonstrated the emission of extreme ultraviolet radiation from thin dielectric films and have investigated the underlying mechanisms.

In 1961, only shortly after the invention of the first laser, scientists exposed silicon dioxide crystals (also known as quartz) to an intense ruby laser to...

Alle Focus-News des Innovations-reports >>>

Anzeige

Anzeige

IHR
JOB & KARRIERE
SERVICE
im innovations-report
in Kooperation mit academics
Veranstaltungen

Potenzial aller Kinder erkennen

29.05.2015 | Veranstaltungen

Cannabis – eine andauernde Kontroverse

29.05.2015 | Veranstaltungen

Frauen können nicht alles haben - Männer aber schon?!

29.05.2015 | Veranstaltungen

 
B2B-VideoLinks
Weitere VideoLinks >>>
Aktuelle Beiträge

Mikhael Subotzky und Patrick Waterhouse erhalten den Deutsche Börse Photography Prize 2015

29.05.2015 | Förderungen Preise

Potenzial aller Kinder erkennen

29.05.2015 | Veranstaltungsnachrichten

HDT - Sommerakademie 2015 für Entwickler und Ingenieure

29.05.2015 | Seminare Workshops