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CSHL researchers solve structure of human protein critical for silencing genes
Date:5/25/2012

proteins from being made and enforcing the silencing of their genes. The discovery of the Argonautes' "slicer" activity stemmed in part from solving the crystal structure of an Argonaute protein from Pyrococcus furiosus, an archebacterium that thrives in extremely high temperatures.

"But we still know nothing about the biological functions or mechanisms of action of archebacterial Argonautes," says Joshua-Tor. "We therefore next focused on solving the structures of Argonautes from higher organisms such as mammals, in which Argonaute functions and target recognition are well documented."

Joshua-Tor's team and other research groups subsequently determined the atomic structures of individual parts of Argonaute proteins from higher organisms. While these studies revealed several important detailsfor example, the interaction between two parts of the Argonaute protein, called the PAZ and Mid domains, with the two ends of guide RNAsJoshua-Tor's goal was to solve the structure of the entire human Argonaute protein in complex with a single human guide RNA.

Overcoming a complicated series of technical challenges, her team has achieved this goal by analyzing the structure of a full-length human Argonaute-2 protein bound to a small RNA called miR-20a, which is known to play a role in cancer development. Although Argonautes from higher organisms diverged from their archebacterial cousins more than three billion years ago, the team's analysis shows remarkable similarity between the two structures, especially in the regions that are important for target recognition and slicing activity.

"Our structure shows that the guide RNA, which is anchored at both ends by the PAZ and Mid domains, kinks and twists its way through the structure of the entire protein, making several points of contact within each domain and with the linker loops that join them," explains Joshua-Tor. "The guide RNA thus acts like a backbone that rigidly locks together the othe
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Contact: Hema Bashyam
bashyam@cshl.edu
516-367-6822
Cold Spring Harbor Laboratory
Source:Eurekalert  

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