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Next-gen reappraisal of interactions within a cancer-associated protein complex
Date:1/15/2014

need to be retested using next-generation approaches. "The old technologies were like observing just one region of the earth from a distant telescope in space and then making assumptions about what the entire earth looked like," Shilatifard says. "With new technologies, we can now see the whole planet."

The methylase in question, named SET1 in yeast and MLL in mammals, is part of a protein aggregate called COMPASS, for COMplex of Proteins ASsociated with Set1. Shilatifard was the first to define the role of COMPASS in chromatin modification. "Over a decade ago, our lab used yeast to show that COMPASS was an H3 methylase," he says. "Since these fundamental systems are highly conserved from yeast to Drosophila to humans, we took advantage of the awesome power of yeast genetics to identify what regulates H3K4 methylation activity."

Part of the paper addresses SET1/MLL regulation by different proteins within yeast COMPASS. Investigators knew that if you lopped off more than half of SET1's front end, levels of DNA-bound trimethylated H3K4 in cells harboring the remaining "stub" were equal to those in cells containing the full-length protein when analyzed in bulk. This led some to presume that the entire front end of SET1/MLL, as well as factors that interact with it, must not be needed to regulate H3K4me3 activity.

The new paper shows that this presumption was not correct. The Shilatifard team first employed biochemical methods to capture every piece of DNA bound to H3K4me3 in the genome of yeast harboring either full-length SET1 or the stub missing the front end. They then sequenced all of those DNA fragments and mapped their position in the yeast genome.

Significantly, they found that even though H3K4me3 levels in bulk were equivalent in normal and mutant cells, H3K4me3 was differentially distributed throughout the genome: in normal cells, H3K4me3 complexes sat primarily on DNA regions that switch adjacent genes on or of
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Contact: Gina Kirchweger
gxk@stowers.org
816-806-1036
Stowers Institute for Medical Research
Source:Eurekalert  

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