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How to build your gate: Decade-old controversy over structure of nuclear pore solved

Date:
July 12, 2013
Source:
European Molecular Biology Laboratory (EMBL)
Summary:
A decade-old controversy over the structure of the nuclear pore has been solved, thanks to a new method which combines thousands of super-resolution microscopy images to reach a precision of less than one nanometer.

At first glance, this may look like the latest view from a telescope, but it was taken by looking not out at the vastness of space, but into a human cell. What could pass for stars or distant galaxies are actually gates to the genome that is locked up inside the cell’s nucleus, as imaged by a super-resolution microscope. By combining images of thousands of these gates, called nuclear pore complexes, scientists at the European Molecular Biology Laboratory (EMBL) in Heidelberg, Germany, have solved a decade-old controversy over how the pieces that form each pore’s ring are arranged.
Credit: EMBL/A. Szymborska

A decade-old controversy over the structure of the nuclear pore has been solved, thanks to a new method which combines thousands of super-resolution microscopy images to reach a precision of less than one nanometre.

It's a parent's nightmare: opening a Lego set and being faced with 500 pieces, but no instructions on how to assemble them into the majestic castle shown on the box. Thanks to a new approach by scientists at the European Molecular Biology Laboratory (EMBL) in Heidelberg, Germany, researchers studying large sets of molecules with vital roles inside our cells can now overcome a similar problem. In a study published online recently in Science, the scientists used super-resolution microscopy to solve a decade-long debate about the structure of the nuclear pore complex, which controls access to the genome by acting as a gate into the cell's nucleus.

Like the flummoxed parent staring at the image on the box, scientists knew the gate's overall shape, from electron tomography studies. And thanks to techniques like X-ray crystallography and single particle electron microscopy, they knew that the ring which studs the nucleus' wall and controls what passes in and out is formed by sixteen or thirty-two copies of a Y-shaped building block. They even knew that each Y is formed by nine proteins. But how the Ys are arranged to form a ring was up for debate.

"When we looked at our images, there was no question: they have to be lying head-to-tail around the hole" says Anna Szymborska, who carried out the work.

To figure out how the Ys were arranged, the EMBL scientists used fluorescent tags to label a series of points along each of the Y's arms and tail, and analysed them under a super-resolution microscope. By combining images from thousands of nuclear pores, they were able to obtain measurements of where each of those points was, in relation to the pore's centre, with a precision of less than a nanometre -- a millionth of a millimetre. The result was a rainbow of rings whose order and spacing meant the Y-shaped molecules in the nuclear pore must lie in an orderly circle around the opening, all with the same arm of the Y pointing toward the pore's centre.

Having resolved this decade-old controversy, the scientists intend to delve deeper into the mysteries of the nuclear pore -- determining whether the circle of Ys is arranged clockwise or anticlockwise, studying it at different stages of assembly, looking at other parts of the pore, and investigating it in three dimensions.

"There's been a lot of interest from other groups," says Jan Ellenberg, who led the work, "so we'll soon be looking into a number of other molecular puzzles, like the different 'machines' that allow a cell to divide, which are also built from hundreds of pieces."

The work was carried out in collaboration with John Briggs' group at EMBL, who helped adapt the image averaging algorithms from electron microscopy to super-resolution microscopy, and Volker Cordes at the Max Planck Institute for Biophysical Chemistry in Gφttingen, Germany, who provided antibodies and advice.


Story Source:

The above story is based on materials provided by European Molecular Biology Laboratory (EMBL). Note: Materials may be edited for content and length.


Journal Reference:

  1. A. Szymborska, A. de Marco, N. Daigle, V. C. Cordes, J. A. G. Briggs, J. Ellenberg. Nuclear Pore Scaffold Structure Analyzed by Super-Resolution Microscopy and Particle Averaging. Science, 2013; DOI: 10.1126/science.1240672

Cite This Page:

European Molecular Biology Laboratory (EMBL). "How to build your gate: Decade-old controversy over structure of nuclear pore solved." ScienceDaily. ScienceDaily, 12 July 2013. <www.sciencedaily.com/releases/2013/07/130712084338.htm>.
European Molecular Biology Laboratory (EMBL). (2013, July 12). How to build your gate: Decade-old controversy over structure of nuclear pore solved. ScienceDaily. Retrieved October 20, 2014 from www.sciencedaily.com/releases/2013/07/130712084338.htm
European Molecular Biology Laboratory (EMBL). "How to build your gate: Decade-old controversy over structure of nuclear pore solved." ScienceDaily. www.sciencedaily.com/releases/2013/07/130712084338.htm (accessed October 20, 2014).

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