Brown scientists map structure of DNA-doctoring protein complex

Dec 06, 2006
DNA dressed for a party
The three-dimensional representation of a DNA-protein complex, developed through fluorescence resonance energy transfer, helped scientists understand the process whereby mobile DNA inserts itself into a chromosome. Image: Marta Radman-Livaja

More than half of the human genome is made up of bits of mobile DNA, which can travel inside the body and insert genes into the chromosomes of target cells. This DNA doctoring not only shapes species over time, it also spreads antibiotic resistance and is used by bacteria that spread Lyme disease and by viruses linked to certain forms of cancer.

Last year in Nature, scientists working in the Brown University lab of Arthur Landy and the Harvard Medical School lab of Thomas Ellenberger announced they had solved the structure of "-integrase ("-Int), the protein "surgeon" that allows mobile DNA to cut into a chromosome, insert its own genes, and then sew the chromosome back up. That work was conducted using the lambda virus, which infects Escherichia coli (E. coli) bacteria and serves as a model that scientists use to understand mobile DNA.

Now scientists in the Landy lab have solved the structure of a DNA-protein complex that acts as a team of "nurses," aiding "-Int during this snip-and-solder procedure known as site-specific recombination. The structure is a three-dimensional representation of the DNA within this complex. Pictured on the cover of the Nov. 17, 2006, journal Molecular Cell, it looks like DNA dressed for a party, a double helix decked with clumps of curly, colorful ribbon. By solving this structure, scientists now know how these six proteins interact with each other and fold DNA during site-specific recombination.

"Once you know how these proteins and DNA are arranged, you have a much better sense of their function," said Xingmin Sun, a postdoctoral research associate in the Landy lab and the lead author of the journal article. "And once you know their function, you begin to see how the real work inside cells gets done."

Sun said solving the structure of the DNA-protein complex called for some creativity. Because it is a string of six proteins, the complex is too big and too flexible to analyze through standard methods such as X-ray crystallography.

Sun used fluorescence resonance energy transfer or FRET, a technique typically used to study small protein complexes in a solution. This time, Sun used FRET to study large protein complexes in a gel. He tagged the DNA with fluorescent dyes and purified the proteins, placing them in a gel that was then shot through with light. Sun measured the wavelengths of light as they bounced between the molecules of dye. Those measurements were then fed into a special software program created by Dale Mierke, a Brown professor of medical science, which plotted their positions to create the structural map.

"The real breakthrough here is successfully using FRET to determine the structure of a large protein-DNA complex," Sun said. "Biologists now have a new tool to help them understand a variety of these complexes, including ones that control cell division, gene expression and DNA replication. So this technique represents a big advance."

Source: Brown University

Explore further: How learning to talk is in the genes

add to favorites email to friend print save as pdf

Related Stories

Unraveling cell division

12 hours ago

CRG researchers shed new light on mitosis. The study published in the Journal of Cell Biology describes how Topo 2 disentangles DNA molecules and is essential for proper cell division

New mechanism in gene regulation revealed

Sep 08, 2014

The information encoded in our genes is translated into proteins, which ultimately mediate biological functions in an organism. Messenger RNA (mRNA) plays an important role, as it is the molecular template used for translation. ...

Plug n' Play protein crystals

Aug 29, 2014

Almost a hundred years ago in 1929 Linus Pauling presented the famous Pauling's Rules to describe the principles governing the structure of complex ionic crystals. These rules essentially describe how the ...

Recommended for you

Gene variant that dramatically reduces 'bad' lipids

10 hours ago

In the first study to emerge from the UK10K Project's cohort of samples from the general public, scientists have identified a rare genetic variant that dramatically reduces levels of certain types of lipids in the blood. ...

New diagnostic method identifies genetic diseases

12 hours ago

People with genetic diseases often have to embark on an odyssey from one doctor to the next. Fewer than half of all patients who are suspected of having a genetic disease actually receive a satisfactory diagnosis. Scientists ...

User comments : 0