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Control of gene expression: Histone occupancy in your genome
Date:4/30/2012

rosstalk." It posits that rather than acting individually, histones within a nucleosome, particularly H3 and H2B, carry on a complex biochemical conversation among themselves based on how they are chemically modified. For example, previous studies had hinted that mono-ubiquitination at one histone (H2B) precedes methylation of another (H3), suggesting that the primary role of the former was to stimulate the latter. Thus, in this paper, Shilatifard and his team asked whether the sole function of H2B mono-ubiquitination was to regulate H3 methylation.

What Jung Shin Lee, Ph.D., a postdoctoral researcher and the study's first author, and Alexander Garrett, a bioinformatics specialist in the Shilatifard lab and co-author, found was unexpected: although yeast carrying mutant Chd1 showed a loss of H2B ubiquitination, H3 methylation was fairly unaffected. "Surprisingly, when we looked genome-wide, we observed that methylation patterns of other histones within nucleosomal complexes were fairly normal in Chd1 mutants," says Lee. "That told us that there are other functions associated with ubiquitination of H2B besides methylation of H3."

To reveal what those functions were, the investigators applied a technique that has been mastered by Penn State biochemist Frank Pugh, Ph.D. The methodology, known as nucleosomal occupancy mapping, can nail down to a single DNA base the very spot on a DNA strand where a complex of histone proteins is sittingnot in just one gene, but in all 5,700 or so yeast genes simultaneously.

Collaborating with Pugh's lab, Shilatifard's group used the technique to map loci where nucleosomes formed in the genome of normal versus Chd1-mutant yeast. Mutants showed decreased occupancy of nucleosomes compared to non-mutant cells, particularly in longer genes, suggesting that mutants have difficulty recreating the proper nucleosome array after gene expression.

"To synthesize RNA from DNA, the normal chromosome structure m
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Contact: Gina Kirchweger
gxk@stowers.org
816-806-1036
Stowers Institute for Medical Research
Source:Eurekalert  

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